Triazolopyridine kinase inhibitors useful for the treatment of degenerative and inflammatory diseases

ABSTRACT

A novel [1,2,4]triazolo[1,5-a]pyridine compound is disclosed that has a formula represented by the following: 
     
       
         
         
             
             
         
       
     
     This compound may be prepared as a pharmaceutical composition, and may be used for the prevention and treatment of a variety of conditions in mammals including humans, including by way of non-limiting example, diseases involving cartilage degradation, bone and/or joint degradation, for example osteoarthritis; and/or conditions involving inflammation or immune responses, such as Crohn&#39;s disease, rheumatoid arthritis, psoriasis, allergic airways disease (e.g. asthma, rhinitis), juvenile idiopathic arthritis, colitis, inflammatory bowel diseases, endotoxin-driven disease states (e.g. complications after bypass surgery or chronic endotoxin states contributing to e.g. chronic cardiac failure), diseases involving impairment of cartilage turnover (e.g. diseases involving the anabolic stimulation of chondrocytes), congenital cartilage malformations, diseases associated with hypersecretion of IL6 and transplantation rejection (e.g. organ transplant rejection) and proliferative diseases.

RELATED APPLICATIONS

The present application claims the benefit under 35 U.S.C. §119 of U.S.Provisional Application No. 61/135,920, filed Jul. 25, 2008, and U.S.Provisional Application No. 61/220,685, filed Jun. 26, 2009, thecontents of each of which are hereby incorporated by reference in theirentireties.

FIELD OF THE INVENTION

The present invention relates to compounds that are inhibitors of JAK, afamily of tyrosine kinases that are involved in the modulation of thedegradation of cartilage, joint degeneration and diseases involving suchdegradation and/or inflammation. The present invention also providesmethods for the production of these compounds, pharmaceuticalcompositions comprising these compounds, methods for the preventionand/or treatment of diseases involving cartilage degradation, boneand/or joint degradation, conditions involving inflammation or immuneresponses, endotoxin-driven disease states, diseases involvingimpairment of cartilage turnover (e.g. diseases involving the anabolicstimulation of chondrocytes), congenital cartilage malformations,diseases associated with hypersecretion of IL6, proliferative diseasesand transplantation rejection (e.g. organ transplant rejection); and/ormethods for the prevention and/or treatment of diseases involvingcartilage degradation, joint degradation and/or inflammation byadministering a compound of the invention.

Janus kinases (JAKs) are cytoplasmic tyrosine kinases that transducecytokine signaling from membrane receptors to STAT transcriptionfactors. Four JAK family members are described, JAK1, JAK2, JAK3 andTYK2. Upon binding of the cytokine to its receptor, JAK family membersauto- and/or transphosphorylate each other, followed by phosphorylationof STATs that then migrate to the nucleus to modulate transcription.JAK-STAT intracellular signal transduction serves the interferons, mostinterleukins, as well as a variety of cytokines and endocrine factorssuch as EPO, TPO, GH, OSM, LIF, CNTF, GM-CSF, PRL Vainchenker W. et al.(2008).

The combination of genetic models and small molecule JAK inhibitorresearch revealed the therapeutic potential of several JAKs. JAK3 isvalidated by mouse and human genetics as an immune-suppression target(O'Shea J. et al. (2004)). JAK3 inhibitors were successfully taken intoclinical development, initially for organ transplant rejection but lateralso in other immuno-inflammatory indications such as rheumatoidarthritis (RA), psoriasis and Crohn's disease(http://clinicaltrials.gov/).

TYK2 is a potential target for immuno-inflammatory diseases, beingvalidated by human genetics and mouse knock-out studies (Levy D. andLoomis C. (2007)).

JAK1 is a novel target in the immuno-inflammatory disease area. JAK1heterodimerizes with the other JAKs to transduce cytokine-drivenpro-inflammatory signaling. Therefore, inhibition of JAK1 and/or otherJAKs is expected to be of therapeutic benefit for a range ofinflammatory conditions as well as for other diseases driven byJAK-mediated signal transduction.

BACKGROUND OF THE INVENTION

Cartilage is an avascular tissue of which chondrocytes are the maincellular component. The chondrocytes in normal articular cartilageoccupy approximately 5% of the tissue volume, while the extra-cellularmatrix makes up the remaining 95% of the tissue. The chondrocytessecrete the components of the matrix, mainly proteoglycans andcollagens, which in turn supply the chondrocytes with an environmentsuitable for their survival under mechanical stress. In cartilage,collagen type II, together with the protein collagen type IX, isarranged in solid fibril-like structures which provide cartilage withgreat mechanical strength. The proteoglycans can absorb water and areresponsible for the resilient and shock absorbing properties of thecartilage.

One of the functional roles of cartilage in the joint is to allow bonesto articulate on each other smoothly. Loss of articular cartilage,therefore, causes the bones to rub against each other leading to painand loss of mobility. The degradation of cartilage can have variouscauses. In inflammatory arthritides, as rheumatoid arthritis forexample, cartilage degradation is caused by the secretion of proteases(e.g. collagenases) by inflamed tissues (the inflamed synovium forexample). Cartilage degradation can also be the result of an injury ofthe cartilage, due to an accident or surgery, or exaggerated loading or‘wear and tear’. The ability of cartilage tissue to regenerate aftersuch insults is limited. Chondrocytes in injured cartilage often displayreduced cartilage synthesizing (anabolic) activity and/or increasedcartilage degrading (catabolic) activity.

The degeneration of cartilage is the hallmark of various diseases, amongwhich rheumatoid arthritis and osteoarthritis are the most prominent.Rheumatoid arthritis (RA) is a chronic joint degenerative disease,characterized by inflammation and destruction of the joint structures.When the disease is unchecked, it leads to substantial disability andpain due to loss of joint functionality and even premature death. Theaim of an RA therapy, therefore, is not only to slow down the diseasebut to attain remission in order to stop the joint destruction. Besidesthe severity of the disease outcome, the high prevalence of RA (˜0.8% ofthe adults are affected worldwide) means a high socio-economic impact.(For reviews on RA, we refer to Smolen and Steiner (2003); Lee andWeinblatt (2001); Choy and Panayi (2001); O'Dell (2004) and Firestein(2003)).

Osteoarthritis (also referred to as OA, or wear-and-tear arthritis) isthe most common form of arthritis and is characterized by loss ofarticular cartilage, often associated with hypertrophy of the bane andpain. The disease mainly affects hands and weight-bearing joints such asknees, hips and spines. This process thins the cartilage. When thesurface area has disappeared due to the thinning, a grade Iosteoarthritis is reached; when the tangential surface area hasdisappeared, grade II osteoarthritis is reached. There are furtherlevels of degeneration and destruction, which affect the deep and thecalcified cartilage layers that border with the subchondral bone. For anextensive review on osteoarthritis, we refer to Wieland et al., 2005.

The clinical manifestations of the development of the osteoarthritiscondition are: increased volume of the joint, pain, crepitation andfunctional disability that lead to pain and reduced mobility of thejoints. When disease further develops, pain at rest emerges. If thecondition persists without correction and/or therapy, the joint isdestroyed leading to disability. Replacement surgery with totalprosthesis is then required.

Therapeutic methods for the correction of the articular cartilagelesions that appear during the osteoarthritic disease have beendeveloped, but so far none of them have been able to mediate theregeneration of articular cartilage in situ and in vivo.

Osteoarthritis is difficult to treat. At present, no cure is availableand treatment focuses on relieving pain and preventing the affectedjoint from becoming deformed. Common treatments include the use ofnon-steroidal anti-inflammatory drugs (NSAIDs). Although dietarysupplements such as chondroitin and glucosamine sulphate have beenadvocated as safe and effective options for the treatment ofosteoarthritis, a recent clinical trial revealed that both treatmentsdid not reduce pain associated to osteoarthritis. (Clegg et al., 2006).Taken together, no disease modifying osteoarthritic drugs are available.

In severe cases, joint replacement may be necessary. This is especiallytrue for hips and knees. If a joint is extremely painful and cannot bereplaced, it may be fused. This procedure stops the pain, but results inthe permanent loss of joint function, making walking and bendingdifficult.

Another possible treatment is the transplantation of cultured autologouschondrocytes. Here, chondral cellular material is taken from thepatient, sent to a laboratory where it is expanded. The material is thenimplanted in the damaged tissues to cover the tissue's defects.

Another treatment includes the intra-articular instillation of Hylan G-F20 (e.g. Synvisc®, Hyalgan®, Artz®), a substance that improvestemporarily the rheology of the synovial fluid, producing an almostimmediate sensation of free movement and a marked reduction of pain.

Other reported methods include application of tendinous, periosteal,fascial, muscular or perichondral grafts; implantation of fibrin orcultured chondrocytes; implantation of synthetic matrices, such ascollagen, carbon fiber; administration of electromagnetic fields. All ofthese have reported minimal and incomplete effects, resulting in a poorquality tissue that can neither support the weighted load nor allow therestoration of an articular function with normal movement.

Stimulation of the anabolic processes, blocking catabolic processes, ora combination of these two, may result in stabilization of thecartilage, and perhaps even reversion of the damage, and thereforeprevent further progression of the disease. Various triggers maystimulate anabolic stimulation of chondrocytes. Insulin-like growthfactor-I (IGF-I) is the predominant anabolic growth factor in synovialfluid and stimulates the synthesis of both proteoglycans and collagen.It has also been shown that members of the bone morphogenetic protein(BMP) family, notably BMP2, BMP4, BMP6, and BMP7, and members of thehuman transforming growth factor-β (TGF-β) family can induce chondrocyteanabolic stimulation (Chubinskaya and Kuettner, 2003). A compound hasrecently been identified that induces anabolic stimulation ofchondrocytes (U.S. Pat. No. 6,500,854; EP 1 391 211). However, most ofthese compounds show severe side effects and, consequently, there is astrong need for compounds that stimulate chondrocyte differentiationwithout these side effects.

Vandeghinste et al. (WO 2005/124342) discovered JAK1 as a target whoseinhibition might have therapeutic relevance for several diseasesincluding OA. JAK1 belongs to the Janus kinase (JAK) family ofcytoplasmic tyrosine kinases, involved in cytokine receptor-mediatedintracellular signal transduction. The JAK family consists of 4 members:JAK1, JAK2, JAK3 and TYK2. JAKs are recruited to cytokine receptors,upon binding of the cytokine, followed by heterodimerization of thecytokine receptor and a shared receptor subunit (common gamma-c chain,gp130). JAKs are then activated by auto- and/or transphosphorylation byanother JAK, resulting in phosphorylation of the receptors andrecruitment and phosphorylation of members of the signal transducer andactivator of transcription (STATs). Phosphorylated STATs dimerize andtranslocate to the nucleus where they bind to enhancer regions ofcytokine-responsive genes. Knockout of the JAK1 gene in micedemonstrated that JAK1 plays essential and nonredundant roles duringdevelopment: JAK1−/− mice died within 24 h after birth and lymphocytedevelopment was severely impaired. Moreover, JAK1−/− cells were not, orless, reactive to cytokines that use class II cytokine receptors,cytokine receptors that use the gamma-c subunit for signaling and thefamily of cytokine receptors that use the gp130 subunit for signaling(Rodig et al., 1998).

Various groups have implicated JAK-STAT signaling in chondrocytebiology. Li et al. (2001) showed that Oncostatin M induces MMP and TIMP3gene expression in primary chondrocytes by activation of JAK/STAT andMAPK signaling pathways. Osaki et al. (2003) showed thatinterferon-gamma mediated inhibition of collagen II in chondrocytesinvolves JAK-STAT signaling. IL1-beta induces cartilage catabolism byreducing the expression of matrix components, and by inducing theexpression of collagenases and inducible nitric oxide synthase (NOS2),which mediates the production of nitric oxide (NO). Otero et al., (2005)showed that leptin and IL1-beta synergistically induced NO production orexpression of NOS2 mRNA in chondrocytes, and that that was blocked by aJAK inhibitor. Legendre et al. (2003) showed that IL6/IL6Receptorinduced downregulation of cartilage-specific matrix genes collagen II,aggrecan core and link protein in bovine articular chondrocytes, andthat this was mediated by JAK/STAT signaling. Therefore, theseobservations suggest a role for JAK kinase activity in cartilagehomeostasis and therapeutic opportunities for JAK kinase inhibitors.

JAK family members have been implicated in additional conditionsincluding myeloproliferative disorders (O'Sullivan et al, 2007, Mol.Immunol. 44(10):2497-506), where mutations in JAK2 have been identified.This indicates that inhibitors of JAK in particular JAK2 may also be ofuse in the treatment of myeloproliferative disorders. Additionally, theJAK family, in particular JAK1, JAK2 and JAK3, has been linked tocancers, in particular leukaemias e.g. acute myeloid leukaemia(O'Sullivan et al, 2007, Mol Immunol. 44(10):2497-506; Xiang et al.,2008, “Identification of somatic JAK1 mutations in patients with acutemyeloid leukemia” Blood First Edition Paper, prepublished online Dec.26, 2007; DOI 10.1182/blood-2007-05-090308) and acute lymphoblasticleukemia (Mullighan et al, 2009) or solid tumours e.g. uterineleiomyosarcoma (Constantinescu et al., 2007, Trends in BiochemicalSciences 33(3): 122-131), prostate cancer (Tam et al., 2007, BritishJournal of Cancer, 97, 378-383) These results indicate that inhibitorsof JAK, in particular of JAK1 and/or JAK2, may also have utility in thetreatment of cancers (leukaemias and solid tumours e.g. uterineleiomyosarcoma, prostate cancer).

In addition, Castleman's disease, multiple myeloma, mesangialproliferative glomerulonephritis, psoriasis, and Kaposi's sarcoma arelikely due to hypersecretion of the cytokine IL-6, whose biologicaleffects are mediated by intracellular JAK-STAT signaling (Tetsuji Naka,Norihiro Nishimoto and Tadamitsu Kishimoto, Arthritis Res 2002, 4 (suppl3):5233-5242). This result shows that inhibitor of JAK, may also findutility in the treatment of said diseases.

A link with autoimmune diseases has been established for JAK3 and Tyk2.Mutations in JAK3 but also in the upstream signaling components gamma-creceptor chain and IL7 receptor account in aggregate for ˜70% of casesof human severe combined immunodeficiency ('OShea et al., 2004). Notethat JAK1 cooperates with JAK3 in transducing signals from the gamma-creceptor chain. Tyk2 polymorphisms are seen in systemic lupuserythematosus (SLE) (O'Sullivan et al, 2007, Mol Immunol.44(10):2497-506). Hence, targeting the JAK family may provide atherapeutic opportunity in the immuno-inflammation area.

The current therapies are not satisfactory and therefore there remains aneed to identify further compounds that may be of use in the treatmentof diseases involving cartilage degradation, bone and/or jointdegradation, for example osteoarthritis; and/or conditions involvinginflammation or immune responses, such as Crohn's disease, rheumatoidarthritis, psoriasis, allergic airways disease (e.g. asthma, rhinitis),juvenile idiopathic arthritis, colitis, inflammatory bowel diseases,endotoxin-driven disease states (e.g. complications after bypass surgeryor chronic endotoxin states contributing to e.g. chronic cardiacfailure), diseases involving impairment of cartilage turnover (e.g.diseases involving the anabolic stimulation of chondrocytes), congenitalcartilage malformations, diseases associated with hypersecretion of IL6and transplantation rejection (e.g. organ transplant rejection).Inhibitors of JAK can also find application in the treatment ofproliferative diseases. In particular the inhibitors of JAK findapplication in the treatment of cancers, especially leukaemias and solidtumours (e.g. uterine leiomyosarcoma, prostate cancer). The presentinvention therefore provides compounds, methods for their manufactureand a pharmaceutical comprising a compound of the invention togetherwith a suitable pharmaceutical carrier. The present invention alsoprovides for the use of a compound of the invention in the preparationof a medicament for the treatment of degenerative joint diseases.

SUMMARY OF THE INVENTION

The present invention is based on the discovery that inhibitors of JAKare useful for the treatment of diseases involving cartilagedegradation, bone and/or joint degradation, for example osteoarthritis;and/or conditions involving inflammation or immune responses, such asCrohn's disease, rheumatoid arthritis, psoriasis, allergic airwaysdisease (e.g. asthma, rhinitis), juvenile idiopathic arthritis, colitis,inflammatory bowel diseases, endotoxin-driven disease states (e.g.complications after bypass surgery or chronic endotoxin statescontributing to e.g. chronic cardiac failure), diseases involvingimpairment of cartilage turnover (e.g. diseases involving the anabolicstimulation of chondrocytes), congenital cartilage malformations,diseases associated with hypersecretion of IL6 and transplantationrejection (e.g. organ transplant rejection). Inhibitors of JAK can alsofind application in the treatment of proliferative diseases. Inparticular the inhibitors of JAK find application in the treatment ofcancers, especially leukaemias and solid tumours (e.g. uterineleiomyosarcoma, prostate cancer). The present invention also providesmethods for the production of these compounds, pharmaceuticalcompositions comprising these compounds and methods for treatingdiseases involving cartilage degradation, joint degradation and/orinflammation by administering a compound of the invention.

Accordingly, in a first aspect of the invention,1,2,4-triazolo[1,5-a]pyridine compounds are disclosed having a Formula(I):

wherein

-   -   Cy1 is selected from aryl and heteroaryl;    -   L1 is selected from a single bond, —O—, —C(O)—, —C[═N(R^(4a))]-,        —N(R^(4a))—, —CON(R^(4a))—, —SO₂N(R^(4a))—, —S(O)₂—,        —N(R^(4a))CO—, or —N(R^(4a))SO₂—;    -   each R¹ is independently selected from C₁-C₆ alkyl, substituted        C₁-C₆ alkyl, acyl, substituted acyl, substituted or        unsubstituted acylamino, substituted or unsubstituted C₁-C₆        alkoxy, substituted or unsubstituted amido, substituted or        unsubstituted amino, substituted sulfinyl, substituted sulfonyl,        substituted or unsubstituted aminosulfonyl, sulfonic acid,        sulfonic acid ester, carboxy, cyano, substituted or        unsubstituted C₃-C₇ cycloalkyl, substituted or unsubstituted 4-7        membered heterocycloalkyl, halo, and hydroxyl;    -   each R^(3a) is independently selected from C₁-C₆ alkyl,        substituted C₁-C₆ alkyl, acyl, substituted acyl, substituted or        unsubstituted acylamino, substituted or unsubstituted C₁-C₆        alkoxy, substituted or unsubstituted amido, alkoxycarbonyl,        substituted alkoxycarbonyl, arylalkyloxy, substituted        arylalkyloxy, substituted or unsubstituted amino, aryl,        substituted aryl, arylalkyl, substituted sulfanyl, substituted        sulfinyl, substituted sulfonyl, substituted or unsubstituted        aminosulfonyl, sulfonic acid, sulfonic acid ester, azido,        carboxy, cyano, substituted or unsubstituted C₃-C₇ cycloalkyl,        substituted or unsubstituted 4-7 membered heterocycloalkyl,        halo, substituted or unsubstituted heteroaryl, hydroxyl, nitro,        and thiol;    -   R^(2a) is selected from substituted or unsubstituted C₁-C₆ alkyl        or substituted or unsubstituted C₃-C₇ cycloalkyl;    -   R^(3b) is independently selected from substituted or        unsubstituted awl, substituted or unsubstituted C₃-C₇        cycloalkyl, substituted or unsubstituted 4-7 membered        heterocycloalkyl, substituted or unsubstituted 5-10 membered        heteroaryl; or R^(3b) is independently selected from O—R^(3c),        CO—R^(3c), and CON(R^(4a))—R^(3c); and R^(3c) is independently        selected from substituted or unsubstituted aryl, substituted or        unsubstituted C₃-C₇ cycloalkyl, substituted or unsubstituted 4-7        membered heterocycloalkyl, substituted or unsubstituted 5-10        membered heteroaryl;    -   each R^(4a), R^(4b) and R^(4c) is independently selected from H,        C₁-C₆ alkyl, substituted C₁-C₆ alkyl, C₃-C₇ cycloalkyl, or        substituted C₃-C₇ cycloalkyl;    -   m1 is 0, 1, or 2; m2 is 0, 1, 2, or 3; and n1 is 0, 1, 2, 3, or        4;    -   provided that        -   when L1 is —O—, —N(R^(4a))—, —CON(R^(4a))—, or            —SO₂N(R^(4a))—, and R^(3b) is other than cycloalkyl,        -   aryl or 5-10 membered heteroaryl, then n1 is 1, 2, 3, or 4;    -   or pharmaceutically acceptable salts or solvates thereof or        solvates of the pharmaceutically acceptable salts.

In a more particular embodiment, the compound is according to FormulaVIa:

-   -   or pharmaceutically acceptable salt or solvate thereof or a        solvate of the pharmaceutically acceptable salt.

In a further aspect, the present invention provides pharmaceuticalcompositions comprising a compound of the invention, and apharmaceutical carrier, excipient or diluent. In this aspect of theinvention, the pharmaceutical composition can comprise one or more ofthe compounds described herein. Moreover, the compounds of the presentinvention useful in the pharmaceutical compositions and treatmentmethods disclosed herein, are all pharmaceutically acceptable asprepared and used.

In a further aspect of the invention, this invention provides a methodof treating a mammal susceptible to or afflicted with a condition fromamong those listed herein, and particularly, such condition as may beassociated with aberrant JAK activity, for example diseases involvingcartilage degradation, bone and/or joint degradation, for exampleosteoarthritis; and/or conditions involving inflammation or immuneresponses, such as Crohn's disease, rheumatoid arthritis, psoriasis,allergic airways disease (e.g. asthma, rhinitis), juvenile idiopathicarthritis, colitis, inflammatory bowel diseases, endotoxin-drivendisease states (e.g. complications after bypass surgery or chronicendotoxin states contributing to e.g. chronic cardiac failure), diseasesinvolving impairment of cartilage turnover (e.g. diseases involving theanabolic stimulation of chondrocytes), congenital cartilagemalformations, diseases associated with hypersecretion of IL6 andtransplantation rejection (e.g. organ transplant rejection). Inhibitorsof JAK can also find application in the treatment of proliferativediseases. In particular the inhibitors of JAK find application in thetreatment of cancers, especially leukaemias and solid tumours (e.g.uterine leiomyosarcoma, prostate cancer). In a particular embodiment thepresent invention provides a method for treating conditions selectedfrom inflammation, such as rheumatoid arthritis, juvenile idiopathicarthritis, psoriasis, allergic airways disease (e.g. asthma, rhinitis),inflammatory bowel diseases (e.g. Crohn's disease, colitis),endotoxin-driven disease states (e.g. complications after bypass surgeryor chronic endotoxin states contributing to e.g. chronic cardiacfailure), and organ transplant rejection; and cartilage, bone and/orjoint degradation or degeneration, such as osteoarthritis, which methodcomprises administering an effective amount of one or more of thepharmaceutical compositions or compounds herein described.

In a further aspect, the present invention provides a method of treatinga mammal susceptible to or afflicted with proliferative disorders inparticular cancer, (e.g. solid tumours), leukaemias, multiple myeloma orpsoriasis, which method comprises administering an effective amount ofone or more of the pharmaceutical compositions or compounds hereindescribed.

In a further aspect, the present invention provides a compound of theinvention for use in the treatment or prevention of a condition selectedfrom those listed herein, particularly such conditions as may beassociated with aberrant JAK activity such as diseases involvingcartilage degradation, bone and/or joint degradation, for exampleosteoarthritis; and/or conditions involving inflammation or immuneresponses, such as Crohn's disease, rheumatoid arthritis, psoriasis,allergic airways disease (e.g. asthma, rhinitis), juvenile idiopathicarthritis, colitis, inflammatory bowel diseases, endotoxin-drivendisease states (e.g. complications after bypass surgery or chronicendotoxin states contributing to e.g. chronic cardiac failure), diseasesinvolving impairment of cartilage turnover (e.g. diseases involving theanabolic stimulation of chondrocytes), congenital cartilagemalformations, diseases associated with hypersecretion of IL6 andtransplantation rejection (e.g. organ transplant rejection). Inhibitorsof JAK can also find application in the treatment of proliferativediseases. In particular the inhibitors of JAK find application in thetreatment of cancers, especially leukaemias and solid tumours (e.g.uterine leiomyosarcoma, prostate cancer). In a specific embodiment, thecondition is selected from inflammation, such as rheumatoid arthritis,juvenile idiopathic arthritis, psoriasis, allergic airways disease (e.g.asthma, rhinitis), inflammatory bowel diseases (e.g. Crohn's disease,colitis), endotoxin-driven disease states (e.g. complications afterbypass surgery or chronic endotoxin states contributing to e.g. chroniccardiac failure), and organ transplant rejection; and cartilage, boneand/or joint degradation or degeneration, such as osteoarthritis.

In a further aspect, the present invention provides a compound of theinvention for use in the treatment or prevention of proliferativedisorders, in particular cancer, (e.g. solid tumours), leukaemias,multiple myeloma or psoriasis.

In yet another method of treatment aspect, this invention provides amethod for treating a mammal susceptible to or afflicted with acondition that is causally related to abnormal JAK activity as describedherein, and comprises administering an effective condition-treating orcondition-preventing amount of one or more of the pharmaceuticalcompositions or compounds herein described.

In a further aspect, the present invention provides a compound of theinvention for use in the treatment or prevention of a condition that iscausally related to abnormal JAK activity.

In additional aspects, this invention provides methods for synthesizingthe compounds of the invention, with representative synthetic protocolsand pathways disclosed later on herein.

Accordingly, it is a principal object of this invention to provide anovel series of compounds, which can modify the activity of JAK and thusprevent or treat any maladies that may be causally related thereto.

It is further an object of this invention to provide a series ofcompounds that can treat or alleviate maladies or symptoms of same, suchas cartilage and/or bone degradation and related inflammation, and jointdiseases, that may be causally related to the activity of JAK.

A still further object of this invention is to provide pharmaceuticalcompositions that may be used in the treatment or prevention of avariety of disease states, including the diseases associated with JAKactivity such as diseases involving cartilage degradation, bone and/orjoint degradation, for example osteoarthritis; and/or conditionsinvolving inflammation or immune responses, such as Crohn's disease,rheumatoid arthritis, psoriasis, allergic airways disease (e.g. asthma,rhinitis), juvenile idiopathic arthritis, colitis, inflammatory boweldiseases, endotoxin-driven disease states (e.g. complications afterbypass surgery or chronic endotoxin states contributing to e.g. chroniccardiac failure), diseases involving impairment of cartilage turnover(e.g. diseases involving the anabolic stimulation of chondrocytes),congenital cartilage malformations, diseases associated withhypersecretion of IL6 and transplantation rejection (e.g. organtransplant rejection). Inhibitors of JAK can also find application inthe treatment of proliferative diseases. In particular the inhibitors ofJAK find application in the treatment of cancers, especially leukaemiasand solid tumours (e.g. uterine leiomyosarcoma, prostate cancer). In aspecific embodiment the condition is selected from inflammation, such asCrohn's disease, rheumatoid arthritis, psoriasis, allergic airwaysdisease (e.g. asthma, rhinitis), juvenile idiopathic arthritis, colitis,inflammatory bowel diseases, endotoxin-driven disease states (e.g.complications after bypass surgery or chronic endotoxin statescontributing to e.g. chronic cardiac failure), and organ transplantrejection; and cartilage, bone and/or joint degradation or degeneration,such as osteoarthritis or cancers (e.g. solid tumours or leukaemias).

Other objects and advantages will become apparent to those skilled inthe art from a consideration of the ensuing detailed description.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The following terms are intended to have the meanings presentedtherewith below and are useful in understanding the description andintended scope of the present invention.

When describing the invention, which may include compounds,pharmaceutical compositions containing such compounds and methods ofusing such compounds and compositions, the following terms, if present,have the following meanings unless otherwise indicated. It should alsobe understood that when described herein any of the moieties definedforth below may be substituted with a variety of substituents, and thatthe respective definitions are intended to include such substitutedmoieties within their scope as set out below. Unless otherwise stated,the term “substituted” is to be defined as set out below. It should befurther understood that the terms “groups” and “radicals” can beconsidered interchangeable when used herein.

The articles “a” and “an” may be used herein to refer to one or to morethan one (i.e. at least one) of the grammatical objects of the article.By way of example “an analogue” means one analogue or more than oneanalogue.

‘Acyl’ or ‘Alkanoyl’ refers to a radical —C(O)R²⁰, where R²⁰ ishydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylmethyl, 4-10membered heterocycloalkyl, aryl, arylalkyl, 5-10 membered heteroaryl orheteroarylalkyl as defined herein. Representative examples include, butare not limited to, formyl, acetyl, cyclohexylcarbonyl,cyclohexylmethylcarbonyl, benzoyl and benzylcarbonyl. Exemplary ‘acyl’groups are —C(O)H, —C(O)—C₁-C₈ alkyl, —C(O)—(CH₂)_(t)(C₆-C₁₀ aryl),—C(O)—(CH₂)_(t)(5-10 membered heteroaryl), —C(O)—(CH₂)_(t)(C₃-C₁₀cycloalkyl), and —C(O)—(CH₂)_(t)(4-10 membered heterocycloalkyl),wherein t is an integer from 0 to 4.

‘Substituted Acyl’ or ‘Substituted Alkanoyl’ refers to a radical—C(O)R²¹, wherein R²¹ is independently

-   -   C₁-C₈ alkyl, substituted with halo or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, each of        which is substituted with unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy.

‘Acylamino’ refers to a radical —NR²²C(O)R²³, where R²² is hydrogen,C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl and R²³ ishydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 memberedheterocycloalkyl, C₆-C₁₀ aryl, arylalkyl, 5-10 membered heteroaryl orheteroarylalkyl, as defined herein. Exemplary ‘acylamino’ include, butare not limited to, formylamino, acetylamino, cyclohexylcarbonylamino,cyclohexylmethyl-carbonylamino, benzoylamino and benzylcarbonylamino.Exemplary ‘acylamino’ groups are —NR^(21′)C(O)—C₁-C₈ alkyl,—NR^(21′)C(O)—(CH₂)_(t)(C₆-C₁₀ aryl), —NR^(21′)C(O)—(CH₂)_(t)(5-10membered heteroaryl), —NR^(20′)C(O)—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—NR^(21′)C(O)—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is aninteger from 0 to 4, each R^(21′) independently represents H or C₁-C₈alkyl.

‘Substituted Acylamino’ refers to a radical —NR²⁴C(O)R²⁵, wherein:

R²⁴ is independently

-   -   H, C₁-C₈ alkyl, substituted with halo or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, each of        which is substituted with unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy; and

R²⁵ is independently

-   -   H, C₁-C₈ alkyl, substituted with halo or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, each of        which is substituted with unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxyl;

provided at least one of R²⁴ and R²⁵ is other than H.

‘Alkoxy’ refers to the group —OR²⁶ where R²⁶ is C₁-C₈ alkyl. Particularalkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,tert-butoxy, sec-butoxy, n-pentoxy, n-henoxy, and 1,2-dimethylbutoxy.Particular alkoxy groups are lower alkoxy, i.e. with between 1 and 6carbon atoms. Further particular alkoxy groups have between 1 and 4carbon atoms.

‘Substituted alkoxy’ refers to an alkoxy group substituted with one ormore of those groups recited in the definition of “substituted” herein,and particularly refers to an alkoxy group having 1 or moresubstituents, for instance from 1 to 5 substituents, and particularlyfrom 1 to 3 substituents, in particular 1 substituent, selected from thegroup consisting of amino, substituted amino, C₆-C₁₀ aryl, -D-aryl,carboxyl, cyano, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl,halogen, 5-10 membered heteroaryl, hydroxyl, nitro, thioalkoxy,thio-O-aryl, thiol, alkyl-S(O)—, aryl-S(O)—, alkyl-S(O)₂— andaryl-S(O)₂—. Exemplary ‘substituted alkoxy’ groups are—O—(CH₂)_(t)(C₆-C₁₀ aryl), —O—(CH₂)_(t)(5-10 membered heteroaryl),—O—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and —O—(CH₂)_(t)(4-10 memberedheterocycloalkyl), wherein t is an integer from 0 to 4 and any aryl,heteroaryl, cycloalkyl or heterocycloalkyl groups present, maythemselves be substituted by unsubstituted C₁-C₄ alkyl, halo,unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl, unsubstitutedC₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy or hydroxy.Particular exemplary ‘substituted alkoxy’ groups are OCF₃, OCH₂CF₃,OCH₂Ph, OCH₂-cyclopropyl, OCH₂CH₂OH, OCH₂CH₂NMe₂.

‘Alkoxycarbonyl’ refers to a radical —C(O)—OR²⁷ where R²⁷ represents anC₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, 4-10 memberedheterocycloalkylalkyl, aralkyl, or 5-10 membered heteroarylalkyl asdefined herein. Exemplary ‘alkoxycarbonyl’ groups are C(O)O—C₁-C₈ alkyl,—C(O)O—(CH₂)_(t)(C₆-C₁₀ aryl), —C(O)O—(CH₂)_(t)(5-10 memberedheteroaryl), —C(O)O—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—C(O)O—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is aninteger from 1 to 4.

‘Substituted Alkoxycarbonyl’ refers to a radical —C(O)—OR²⁸ where R²⁸represents:

-   -   C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ cycloalkylalkyl, or 4-10        membered heterocycloalkylalkyl, each of which is substituted        with halo, substituted or unsubstituted amino, or hydroxy; or    -   C₆-C₁₀ aralkyl, or 5-10 membered heteroarylalkyl, each of which        is substituted with unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxyl.

‘Alkyl’ means straight or branched aliphatic hydrocarbon having 1 to 20carbon atoms. Particular alkyl has 1 to 12 carbon atoms. More particularis lower alkyl which has 1 to 6 carbon atoms. A further particular grouphas 1 to 4 carbon atoms. Exemplary straight chained groups includemethyl, ethyl n-propyl, and n-butyl. Branched means that one or morelower alkyl groups such as methyl, ethyl, propyl or butyl is attached toa linear alkyl chain, exemplary branched chain groups include isopropyl,iso-butyl, t-butyl and isoamyl.

‘Substituted alkyl’ refers to an alkyl group as defined abovesubstituted with one or more of those groups recited in the definitionof “substituted” herein, and particularly refers to an alkyl grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, in particular 1 substituent,selected from the group consisting of acyl, acylamino, acyloxy (—O-acylor —OC(O)R²⁰), alkoxy, alkoxycarbonyl, alkoxycarbonylamino(—NR″-alkoxycarbonyl or —NH—C(O)—OR²⁷), amino, substituted amino,aminocarbonyl (carbamoyl or amido or —C(O)—NR″₂), aminocarbonylamino(—NR″—C(O)—NR″₂), aminocarbonyloxy (—O—C(O)—NR″₂), aminosulfonyl,sulfonylamino, aryl, —O-aryl, azido, carboxyl, cyano, cycloalkyl,halogen, hydroxy, heteroaryl, nitro, thiol, —S-alkyl, —S-aryl,—S(O)-alkyl, —S(O)-aryl, —S(O)₂-alkyl, and —S(O)₂-aryl. In a particularembodiment ‘substituted alkyl’ refers to a C₁-C₈ alkyl group substitutedwith halo, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido,—NR′″SO₂R″, —SO₂NR″R′″, —C(O)R″, —C(O)OR″, —OC(O)R″, —NRC′″(O)R″,—C(O)NR″R′″, —NR″R′″, or —(CR′″R″″)_(m)OR′″; wherein each R″ isindependently selected from H, C₁-C₈ alkyl, —(CH₂)_(t)(C₆-C₁₀ aryl),—(CH₂)_(t)(5-10 membered heteroaryl), —(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is an integer from0 to 4 and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl groupspresent, may themselves be substituted by unsubstituted alkyl, halo,unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl, unsubstitutedC₁-C₄ hydroxyalkyl, or unsubstituted haloalkoxy or hydroxy. Each of R′″and R″″ independently represents H or C₁-C₈ alkyl.

‘Amino’ refers to the radical —NH₂.

‘Substituted amino’ refers to an amino group substituted with one ormore of those groups recited in the definition of ‘substituted’ herein,and particularly refers to the group —N(R³³)₂ where each R³³ isindependently selected from:

-   -   hydrogen, C₁-C₈ alkyl, C₆-C₁₀ aryl, 5-10 membered heteroaryl,        4-10 membered heterocycloalkyl, or C₃-C₁₀ cycloalkyl; or    -   C₁-C₈ alkyl, substituted with halo or hydroxy; or    -   —(CH₂)_(t)(C₆-C₁₀ aryl), —(CH₂)_(t)(5-10 membered heteroaryl),        —(CH₂)_(t)(C₃-C₁₀ cycloalkyl) or —(CH₂)_(t)(4-10 membered        heterocycloalkyl) wherein t is an integer between 0 and 8, each        of which is substituted by unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy; or    -   both R³³ groups are joined to form an alkylene group.        When both R³³ groups are hydrogen, —N(R³³)₂ is an amino group.        Exemplary ‘substituted amino’ groups are —NR^(33′) —C₁-C₈ alkyl,        —NR^(33′)—(CH₂)_(t)(C₆-C₁₀ aryl), —NR^(33′)—(CH₂)_(t)(5-10        membered heteroaryl), —NR^(33′)—(CH₂)_(t)(C₃-C₁₀ cyclo alkyl),        and —NR^(33′)'(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein        t is an integer from 0 to 4, each R^(33′)independently        represents H or C₁-C₈ alkyl; and any alkyl groups present, may        themselves be substituted by halo, substituted or unsubstituted        amino, or hydroxy; and any aryl, heteroaryl, cycloalkyl or        heterocycloalkyl groups present, may themselves be substituted        by unsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,        unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl,        or unsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Aminosulfonyl’ or ‘Sulfonamide’ refers to the radical —S(O₂)NH₂.

‘Substituted aminosulfonyl’ or ‘substituted sulfonamide’ refers to aradical such as —S(O₂)N(R⁴⁸)₂ wherein each R⁴⁸ is independently selectedfrom:

-   -   H, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered        heterocycloalkyl, C₆-C₁₀ aryl, aralkyl, 5-10 membered        heteroaryl, and heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, substituted        by unsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,        unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl,        or unsubstituted C₁-C₄ haloalkoxy or hydroxy;

provided that at least one R⁴⁸ is other than H.

Exemplary ‘substituted aminosulfonyl’ or ‘substituted sulfonamide’groups are —S(O₂)N(R^(48′))—C₁-C₈ alkyl,—S(O₂)N(R^(48′))—(CH₂)_(t)(C₆-C₁₀ aryl), —S(O₂)N(R^(48′))—(CH₂)_(t)(5-10membered heteroaryl), —S(O₂)N(R^(48′))—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—S(O₂)N(R^(48′))—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t isan integer from 0 to 4; each R^(48′) independently represents H or C₁-C₈alkyl; and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl groupspresent, may themselves be substituted by unsubstituted C₁-C₄ alkyl,halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy orhydroxy.

‘Aralkyl’ or ‘arylalkyl’ refers to an alkyl group, as defined above,substituted with one or more aryl groups, as defined above. Particulararalkyl or arylalkyl groups are alkyl groups substituted with one arylgroup.

‘Substituted Aralkyl’ or ‘substituted arylalkyl’ refers to an alkylgroup, as defined above, substituted with one or more aryl groups; andat least one of any aryl group present, may themselves be substituted byunsubstituted C₁-C₄ alkyl, halo, cyano, unsubstituted C₁-C₄ alkoxy,unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, orunsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Aryl’ refers to a monovalent aromatic hydrocarbon group derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. In particular aryl refers to an aromatic ringstructure, mono-cyclic or poly-cyclic that includes from 5 to 12 ringmembers, more usually 6 to 10. Where the aryl group is a monocyclic ringsystem it preferentially contains 6 carbon atoms. Typical aryl groupsinclude, but are not limited to, groups derived from aceanthrylene,acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene,hexylene, as-indacene, s-indacene, indane, indene, naphthalene,octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene,pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,pleiadene, pyrene, pyranthrene, rubicene, triphenylene andtrinaphthalene. Particularly aryl groups include phenyl, naphthyl,indenyl, and tetrahydronaphthyl.

‘Substituted Aryl’ refers to an aryl group substituted with one or moreof those groups recited in the definition of ‘substituted’ herein, andparticularly refers to an aryl group that may optionally be substitutedwith 1 or more substituents, for instance from 1 to 5 substituents,particularly 1 to 3 substituents, in particular 1 substituent.Particularly, ‘Substituted Aryl’ refers to an aryl group substitutedwith one or more of groups selected from halo, C₁-C₈ alkyl, C₁-C₈haloalkyl, C₁-C₈ haloalkoxy, cyano, hydroxy, C₁-C₈ alkoxy, and amino.

Examples of representative substituted aryls include the following

In these formulae one of R⁴⁹ and R⁵⁰ may be hydrogen and at least one ofR⁴⁹ and R⁵⁰ is each independently selected from C₁-C₈ alkyl, 4-10membered heterocycloalkyl, alkanoyl, C₁-C₈ alkoxy, hetero-O-aryl,alkylamino, arylamino, heteroarylamino, NR⁵¹COR⁵², NR⁵¹SOR⁵²NR⁵¹SO₂NR⁵², COOalkyl, COOaryl, CONR⁵¹R⁵², CONR⁵¹OR⁵², NR⁵¹R⁵²,SO₂NR⁵¹R⁵², S-alkyl, SOalkyl, SO₂-alkyl, Saryl, SOaryl, SO₂-aryl; or R⁴⁹and R⁵⁰ may be joined to form a cyclic ring (saturated or unsaturated)from 5 to 8 atoms, optionally containing one or more heteroatomsselected from the group N, O or S. R⁵¹, and R⁵² are independentlyhydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, C₃-C₁₀ cycloalkyl, 4-10 memberedheterocycloalkyl, C₆-C₁₀ aryl, substituted aryl, 5-10 memberedheteroaryl.

‘Arylalkyloxy’ refers to an —O-alkylaryl radical where alkylaryl is asdefined herein.

‘Substituted Arylalkyloxy’ refers to an —O-alkylaryl radical wherealkylaryl is as defined herein; and any aryl groups present, maythemselves be substituted by unsubstituted C₁-C₄ alkyl, halo, cyano,unsubstituted C₁-C₄ alkoxy, unsubstituted C₁₋₄ haloalkyl, unsubstitutedC₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Azido’ refers to the radical —N₃.

‘Carbamoyl or amido’ refers to the radical —C(O)NH₂.

‘Substituted Carbamoyl or substituted amido’ refers to the radical—C(O)N(R⁵³)₂ wherein each R⁵³ is independently

-   -   H, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered        heterocycloalkyl, C₆-C₁₀ aryl, aralkyl, 5-10 membered        heteroaryl, and heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, each of        which is substituted by unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy;

provided that at least one R⁵³ is other than H.

Exemplary ‘Substituted Amido/Carbamoyl’ groups are —C(O)NR^(53′)—C₁-C₈alkyl, —C(O)NR^(53′)—(CH₂)_(t)(C₆-C₁₀ aryl), —C(O)N^(53′)—(CH₂)_(t)(5-10membered heteroaryl), —C(O)NR^(53′)—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—C(O)NR^(53′)—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is aninteger from 0 to 4, each R^(53′) independently represents H or C₁-C₈alkyl and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl groupspresent, may themselves be substituted by unsubstituted C₁-C₄ alkyl,halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy orhydroxy.

‘Carboxy’ refers to the radical —C(O)OH.

‘Cycloalkyl’ refers to cyclic non-aromatic hydrocarbyl groups havingfrom 3 to 10 carbon atoms. Such cycloalkyl groups include, by way ofexample, single ring structures such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

‘Substituted cycloalkyl’ refers to a cycloalkyl group as defined abovesubstituted with one or more of those groups recited in the definitionof ‘substituted’ herein, and particularly refers to a cycloalkyl grouphaving 1 or more substituents, for instance from 1 to 5 substituents,and particularly from 1 to 3 substituents, in particular 1 substituent.

‘Cyano’ refers to the radical —CN.

‘Halo’ or ‘halogen’ refers to fluoro (F), chloro (Cl), bromo (Br) andiodo (I). Particular halo groups are either fluoro or chloro.

‘Hetero’ when used to describe a compound or a group present on acompound means that one or more carbon atoms in the compound or grouphave been replaced by a nitrogen, oxygen, or sulfur heteroatom. Heteromay be applied to any of the hydrocarbyl groups described above such asalkyl, e.g. heteroalkyl, cycloalkyl, e.g. heterocycloalkyl, aryl, e.g.heteroaryl, cycloalkenyl, e.g. cycloheteroalkenyl, and the like havingfrom 1 to 5, and particularly from 1 to 3 heteroatoms.

‘Heteroaryl’ means an aromatic ring structure, mono-cyclic orpolycyclic, that includes one or more heteroatoms and 5 to 12 ringmembers, more usually 5 to 10 ring members. The heteroaryl group can be,for example, a five membered or six membered monocyclic ring or abicyclic structure formed from fused five and six membered rings or twofused six membered rings or, by way of a further example, two fused fivemembered rings. Each ring may contain up to four heteroatoms typicallyselected from nitrogen, sulphur and oxygen. Typically the heteroarylring will contain up to 4 heteroatoms, more typically up to 3heteroatoms, more usually up to 2, for example a single heteroatom. Inone embodiment, the heteroaryl ring contains at least one ring nitrogenatom. The nitrogen atoms in the heteroaryl rings can be basic, as in thecase of an imidazole or pyridine, or essentially non-basic as in thecase of an indole or pyrrole nitrogen. In general the number of basicnitrogen atoms present in the heteroaryl group, including any aminogroup substituents of the ring, will be less than five. Examples of fivemembered monocyclic heteroaryl groups include but are not limited topyrrole, furan, thiophene, imidazole, furazan, oxazole, oxadiazole,oxatriazole, isoxazole, thiazole, isothiazole, pyrazole, triazole andtetrazole groups. Examples of six membered monocyclic heteroaryl groupsinclude but are not limited to pyridine, pyrazine, pyridazine,pyrimidine and triazine. Particular examples of bicyclic heteroarylgroups containing a five membered ring fused to another five memberedring include but are not limited to imidazothiazole andimidazoimidazole. Particular examples of bicyclic heteroaryl groupscontaining a six membered ring fused to a five membered ring include butare not limited to benzfuran, benzthiophene, benzimidazole, benzoxazole,isobenzoxazole, benzisoxazole, benzthiazole, benzisothiazole,isobenzofuran, indole, isoindole, isoindolone, indolizine, indoline,isoindoline, purine (e.g., adenine, guanine), indazole,pyrazolopyrimidine, triazolopyrimidine, benzodioxole andpyrazolopyridine groups. Particular examples of bicyclic heteroarylgroups containing two fused six membered rings include but are notlimited to quinoline, isoquinoline, chroman, thiochroman, chromene,isochromene, chroman, isochroman, benzodioxan, quinolizinc, benzoxazine,benzodiazinc, pyridopyridine, quinoxaline, quinazoline, cinnoline,phthalazine, naphthyridine and pteridine groups. Particular heteroarylgroups are those derived from thiophene, pyrrole, benzothiophene,benzofuran, indole, pyridine, quinoline, imidazole, oxazole andpyrazine.

Examples of representative aryl having hetero atoms containingsubstitution include the following:

wherein each W is selected from C(R⁵⁴)₂, NR⁵⁴, O and S; and each Y isselected from carbonyl, NR⁵⁴, O and S; and R⁵⁴ is independentlyhydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 memberedheterocycloalkyl, C₆-C₁₀ aryl, and 5-10 membered heteroaryl.

Examples of representative heteroaryls include the following:

wherein each Y is selected from carbonyl, N, NR⁵⁵, O and S; and R⁵⁵ isindependently hydrogen, C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 memberedheterocycloalkyl, C₆-C₁₀ aryl, and 5-10 membered heteroaryl.

As used herein, the term ‘heterocycloalkyl’ refers to a 4-10 membered,stable heterocyclic non-aromatic ring and/or including rings containingone or more heteroatoms independently selected from N, O and S, fusedthereto. A fused heterocyclic ring system may include carbocyclic ringsand need only include one heterocyclic ring. Examples of heterocyclicrings include, but are not limited to, morpholine, piperidine (e.g.1-piperidinyl, 2-piperidinyl, 3-piperidinyl and 4-piperidinyl),pyrrolidine (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl),pyrrolidone, pyran (2H-pyran or 4H-pyran), dihydrothiophene,dihydropyran, dihydrofuran, dihydrothiazole, tetrahydrofuran,tetrahydrothiophene, dioxane, tetrahydropyran (e.g. 4-tetrahydropyranyl), imidazoline, imidazolidinone, oxazoline, thiazoline,2-pyrazoline, pyrazolidine, piperazine, and N-alkyl piperazines such asN-methyl piperazine. Further examples include thiomorpholine and itsS-oxide and S,S-dioxide (particularly thiomorpholine). Still furtherexamples include azetidine, piperidone, piperazone, and N-alkylpiperidines such as N-methyl piperidine. Particular examples ofheterocycloalkyl groups are shown in the following illustrativeexamples:

wherein each W is selected from CR⁵⁶, C(R⁵⁶)₂, NR⁵⁶, O and S; and each Yis selected from NR⁵⁶, O and S; and R⁵⁶ is independently hydrogen, C₁-C₈alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,5-10 membered heteroaryl, These heterocycloalkyl rings may be optionallysubstituted with one or more groups selected from the group consistingof acyl, acylamino, acyloxy (—O-acyl or —OC(O)R²⁰), alkoxy,alkoxycarbonyl, alkoxycarbonylamino (—NR″-alkoxycarbonyl or—NH—C(O)—OR²⁷), amino, substituted amino, aminocarbonyl (amido or—C(O)—NR″₂), aminocarbonylamino (—NR″-C(O)—NR″₂), aminocarbonyloxy(—O—C(O)—NR″₂), aminosulfonyl, sulfonylamino, aryl, —O-aryl, azido,carboxyl, cyano, cycloalkyl, halogen, hydroxy, nitro, thiol, —S-alkyl,—S-aryl, —S(O)-alkyl, —S(O)-aryl, —S(O)₂-alkyl, and —S(O)₂-aryl.Substituting groups include carbonyl or thiocarbonyl which provide, forexample, lactam and urea derivatives.

‘Hydroxy’ refers to the radical —OH.

‘Nitro’ refers to the radical —NO₂.

‘Substituted’ refers to a group in which one or more hydrogen atoms areeach independently replaced with the same or different substituent(s).Typical substituents may be selected from the group consisting of:

halogen, —R⁵⁷, —O⁻, ═O, —OR⁵⁷, —SR⁵⁷, —S⁻, ═S, NR⁵⁷R⁵⁸, ═NR⁵⁷, —CCl₃,—CF₃, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —S(O)₂O⁻, —S(O)₂OH,—S(O)₂R⁵⁷, —OS(O₂)O⁻, —OS(O)₂R⁵⁷, —P(O)(O⁻)₂, —P(O)(OR⁵⁷)(O⁻),—OP(O)(OR⁵⁷)(OR⁵⁸), —C(O)R⁵⁷, —C(S)R⁵⁷, —C(O)OR⁵⁷, —C(O)NR⁵⁷R⁵⁸,—C(O)O⁻, —C(S)OR⁵⁷, —NR⁵⁹C(O)NR⁵⁷R⁵⁸, —NR⁵⁹C(S)NR⁵⁷R⁵⁸,—NR⁶⁰C(NR⁵⁹)NR⁵⁷R⁵⁸ and —C(NR⁵⁹)NR⁵⁷R⁵⁸;

wherein each R⁵⁷, R⁵⁸, R⁵⁹ and R⁶⁰ are independently:

-   -   hydrogen, C₁-C₈ alkyl, C₆-C₁₀ aryl, arylalkyl, C₃-C₁₀        cycloalkyl, 4-10 membered heterocycloalkyl, 5-10 membered        heteroaryl, heteroarylalkyl; or    -   C₁-C₈ alkyl substituted with halo or hydroxy; or    -   C₆-C₁₀ aryl, 5-10 membered heteroaryl, C₆-C₁₀ cycloalkyl or 4-10        membered heterocycloalkyl substituted by unsubstituted C₁-C₄        alkyl, halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄        haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted        C₁-C₄ haloalkoxy or hydroxy.        In a particular embodiment, substituted groups are substituted        with one or more substituents, particularly with 1 to 3        substituents, in particular with one substituent group.

In a further particular embodiment the substituent group or groups areselected from: halo, cyano, nitro, trifluoromethyl, trifluoromethoxy,azido, —NR′″SO₂R″, —SO₂NR″R′″, —C(O)R″, —C(O)OR″, —OC(O)R″, —NR′″C(O)R″,—C(O)NR″R′″, —(CR′″R′″)_(m)OR′″, wherein, each R″ is independentlyselected from H, C₁-C₈ alkyl, —(CH₂)_(t)(C₆-C₁₀ aryl), —(CH₂)_(t)(5-10membered heteroaryl), —(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and —(CH₂)_(t)(4-10membered heterocycloalkyl), wherein t is an integer from 0 to 4; and

-   -   any alkyl groups present, may themselves be substituted by halo        or hydroxy; and    -   any aryl, heteroaryl, cycloalkyl or heterocycloalkyl groups        present, may themselves be substituted by unsubstituted C₁-C₄        alkyl, halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄        haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted        C₁-C₄ haloalkoxy or hydroxy. Each R″ independently represents H        or C₁-C₆alkyl.

‘Substituted sulfanyl’ refers to the group —SR⁶¹, wherein R⁶¹ isselected from:

-   -   C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl,        C₆-C₁₀ aryl, aralkyl, 5-10 membered heteroaryl, and        heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo, substituted or unsubstituted        amino, or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, each of        which is substituted by unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy.

Exemplary ‘substituted sulfanyl’ groups are —S—(C₁-C₈ alkyl) and—S—(C₃-C₁₀ cycloalkyl), —S—(CH₂)_(t)(C₆-C₁₀ aryl), —S—(CH₂)_(t)(5-10membered heteroaryl), —S—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—S—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is an integerfrom 0 to 4 and any aryl, heteroaryl, cycloalkyl or heterocycloalkylgroups present, may themselves be substituted by unsubstituted C₁-C₄alkyl, halo, unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄ haloalkoxy orhydroxy.

‘Substituted sulfinyl’ refers to the group —S(O)R⁶⁸, wherein R⁶⁸ isselected from:

-   -   C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl,        C₆-C₁₀ aryl, aralkyl, 5-10 membered heteroaryl, and        heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo, substituted or unsubstituted        amino, or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, substituted        by unsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,        unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl,        or unsubstituted C₁-C₄ haloalkoxy or hydroxy.

Exemplary ‘substituted sulfinyl’ groups are —S(O)—(C₁-C₈ alkyl) and—S(O)—(C₃-C₁₀ cycloalkyl), —S(O)—(CH₂)_(t)(C₆-C₁₀ aryl), —S(O)—(CH₂)_(t)(5-10 membered heteroaryl), —S(O)—(CH₂)_(t)(C₃-C₁₀ cycloalkyl),and —S(O)—(C₁₋₁₂)_(t)(4-10 membered heterocycloalkyl), wherein t is aninteger from 0 to 4 and any aryl, heteroaryl, cycloalkyl orheterocycloalkyl groups present, may themselves be substituted byunsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, orunsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Substituted sulfonyl’ refers to the group —S(O)₂R⁷⁵, wherein R⁷⁵ isselected from:

-   -   C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl,        C₆-C₁₀ aryl, aralkyl, 5-10 membered heteroaryl, and        heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo, substituted or unsubstituted        amino, or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, each of        which is substituted by unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy.

Exemplary ‘substituted sulfonyl’ groups are —S(O)₂—(C₁-C₈ alkyl) and—S(O)₂—(C₃-C₁₀ cycloalkyl), —S(O)₂—(CH₂)_(t)(C₆-C₁₀ aryl),—S(O)₂—(CH₂)_(t)(5-10 membered heteroaryl), —S(O)₂—(CH₂)_(t)(C₃-C₁₀cycloalkyl), and —S(O)₂—(CH₂)_(t)(4-10 membered heterocycloalkyl),wherein t is an integer from 0 to 4 and any aryl, heteroaryl, cycloalkylor heterocycloalkyl groups present, may themselves be substituted byunsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, orunsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Sulfo’ or ‘sulfonic acid’ refers to a radical such as —SO₃H.

‘Substituted sulfo’ or ‘sulfonic acid ester’ refers to the group—S(O)₂OR⁸², wherein R⁸² is selected from:

-   -   C₁-C₈ alkyl, C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl,        C₆-C₁₀ aryl, aralkyl, 5-10 membered heteroaryl, and        heteroaralkyl; or    -   C₁-C₈ alkyl substituted with halo, substituted or unsubstituted        amino, or hydroxy; or    -   C₃-C₁₀ cycloalkyl, 4-10 membered heterocycloalkyl, C₆-C₁₀ aryl,        aralkyl, 5-10 membered heteroaryl, or heteroaralkyl, each of        which is substituted by unsubstituted C₁-C₄ alkyl, halo,        unsubstituted C₁-C₄ alkoxy, unsubstituted C₁-C₄ haloalkyl,        unsubstituted C₁-C₄ hydroxyalkyl, or unsubstituted C₁-C₄        haloalkoxy or hydroxy.

Exemplary ‘Substituted sulfo’ or ‘sulfonic acid ester’ groups are—S(O)₂—O—(C₁-C₈ alkyl) and —S(O)₂—O—(C₃-C₁₀ cycloalkyl),—S(O)₂—O—(CH₂)_(t)(C₆-C₁₀ aryl), —S(O)₂—O—(CH₂)_(t)(5-10 memberedheteroaryl), —S(O)₂—O—(CH₂)_(t)(C₃-C₁₀ cycloalkyl), and—S(O)₂—O—(CH₂)_(t)(4-10 membered heterocycloalkyl), wherein t is aninteger from 0 to 4 and any aryl, heteroaryl, cycloalkyl orheterocycloalkyl groups present, may themselves be substituted byunsubstituted C₁-C₄ alkyl, halo, unsubstituted C₁-C₄ alkoxy,unsubstituted C₁-C₄ haloalkyl, unsubstituted C₁-C₄ hydroxyalkyl, orunsubstituted C₁-C₄ haloalkoxy or hydroxy.

‘Thiol’ refers to the group —SH.

One having ordinary skill in the art of organic synthesis will recognizethat the maximum number of heteroatoms in a stable, chemically feasibleheterocyclic ring, whether it is aromatic or non aromatic, is determinedby the size of the ring, the degree of unsaturation and the valence ofthe heteroatoms. In general, a heterocyclic ring may have one to fourheteroatoms so long as the heteroaromatic ring is chemically feasibleand stable.

‘Pharmaceutically acceptable’ means approved or approvable by aregulatory agency of the Federal or a state government or thecorresponding agency in countries other than the United States, or thatis listed in the U.S. Pharmacopoeia or other generally recognizedpharmacopoeia for use in animals, and more particularly, in humans.

‘Pharmaceutically acceptable salt’ refers to a salt of a compound of theinvention that is pharmaceutically acceptable and that possesses thedesired pharmacological activity of the parent compound. In particular,such salts are non-toxic may be inorganic or organic acid addition saltsand base addition salts. Specifically, such salts include: (1) acidaddition salts, formed with inorganic acids such as hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike; or formed with organic acids such as acetic acid, propionic acid,hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid,lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, N-methylglucamine and thelike. Salts further include, by way of example only, sodium, potassium,calcium, magnesium, ammonium, tetraalkylammonium, and the like; and whenthe compound contains a basic functionality, salts of non toxic organicor inorganic acids, such as hydrochloride, hydrobromide, tartrate,mesylate, acetate, maleate, oxalate and the like. The term“pharmaceutically acceptable cation” refers to an acceptable cationiccounter-ion of an acidic functional group. Such cations are exemplifiedby sodium, potassium, calcium, magnesium, ammonium, tetraalkylammoniumcations, and the like.

‘Pharmaceutically acceptable vehicle’ refers to a diluent, adjuvant,excipient or carrier with which a compound of the invention isadministered.

‘Prodrugs’ refers to compounds, including derivatives of the compoundsof the invention, which have cleavable groups and become by solvolysisor under physiological conditions the compounds of the invention whichare pharmaceutically active in vivo. Such examples include, but are notlimited to, choline ester derivatives and the like, N-alkylmorpholineesters and the like.

‘Solvate’ refers to forms of the compound that are associated with asolvent, usually by a solvolysis reaction. This physical associationincludes hydrogen bonding. Conventional solvents include water, ethanol,acetic acid and the like. The compounds of the invention may be preparede.g. in crystalline form and may be solvated or hydrated. Suitablesolvates include pharmaceutically acceptable solvates, such as hydrates,and further include both stoichiometric solvates and non-stoichiometricsolvates. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. ‘Solvate’ encompasses bothsolution-phase and insoluble solvates. Representative solvates includehydrates, ethanolates and methanolates.

‘Subject’ includes humans. The terms ‘human’, ‘patient’ and ‘subject’are used interchangeably herein.

‘Therapeutically effective amount’ means the amount of a compound that,when administered to a subject for treating a disease, is sufficient toeffect such treatment for the disease. The “therapeutically effectiveamount” can vary depending on the compound, the disease and itsseverity, and the age, weight, etc., of the subject to be treated.

‘Preventing’ or ‘prevention’ refers to a reduction in risk of acquiringor developing a disease or disorder (i.e., causing at least one of theclinical symptoms of the disease not to develop in a subject that may beexposed to a disease-causing agent, or predisposed to the disease inadvance of disease onset.

The term ‘prophylaxis’ is related to ‘prevention’, and refers to ameasure or procedure the purpose of which is to prevent, rather than totreat or cure a disease. Non-limiting examples of prophylactic measuresmay include the administration of vaccines; the administration of lowmolecular weight heparin to hospital patients at risk for thrombosisdue, for example, to immobilization; and the administration of ananti-malarial agent such as chloroquine, in advance of a visit to ageographical region where malaria is endemic or the risk of contractingmalaria is high.

‘Treating’ or ‘treatment’ of any disease or disorder refers, in oneembodiment, to ameliorating the disease or disorder (i.e., arresting thedisease or reducing the manifestation, extent or severity of at leastone of the clinical symptoms thereof). In another embodiment ‘treating’or ‘treatment’ refers to ameliorating at least one physical parameter,which may not be discernible by the subject. In yet another embodiment,‘treating’ or ‘treatment’ refers to modulating the disease or disorder,either physically, (e.g., stabilization of a discernible symptom),physiologically, (e.g., stabilization of a physical parameter), or both.In a further embodiment, ‘treating’ or ‘treatment’ relates to slowingthe progression of the disease.

As used herein the term ‘condition(s) involving inflammation’ refers tothe group of conditions including, rheumatoid arthritis, osteoarthritis,juvenile idiopathic arthritis, psoriasis, allergic airway disease (e.g.asthma, rhinitis), inflammatory bowel diseases (e.g. Crohn's disease,colitis), endotoxin-driven disease states (e.g. complications afterbypass surgery or chronic endotoxin states contributing to e.g. chroniccardiac failure), and related diseases involving cartilage, such as thatof the joints. Particularly the term refers to rheumatoid arthritis,osteoarthritis, allergic airway disease (e.g. asthma) and inflammatorybowel diseases.

As used herein the term ‘condition(s) involving an immune response’ or‘autoimmune diseases’ are used interchangeably and refer to refers tothe group of diseases including obstructive airways disease, includingconditions such as COPD, asthma (e.g. intrinsic asthma, extrinsicasthma, dust asthma, infantily asthma) particularly chronic orinveterate asthma (for example late asthma and airwayhyperreponsiveness), bronchitis, including bronchial asthma, systemiclupus erythematosus (SLE), multiple sclerosis, type I diabetes mellitusand complications associated therewith, atopic eczema (atopicdermatitis), contact dermatitis and further eczematous dermatitis,inflammatory bowel disease (e.g. Crohn's disease and ulcerativecolitis), atherosclerosis and amyotrophic lateral sclerosis.Particularly the term refers to COPD, asthma, systemic lupuserythematosis, type I diabetes mellitus and inflammatory bowel disease.

As used herein the term ‘transplantation rejection’ refers to the acuteor chronic rejection of cells, tissue or solid organ allo- or xenograftsof e.g. pancreatic islets, stem cells, bone marrow, skin, muscle,corneal tissue, neuronal tissue, heart, lung, combined heart-lung,kidney, liver, bowel, pancreas, trachea or esophagus, orgraft-versus-host diseases.

As used herein the term ‘proliferative disease(s)’ refers to conditionssuch as cancer (e.g. uterine leiomyosarcoma or prostate cancer),myeloproliferative disorders (e.g. polycythemia vera, essentialthrombocytosis and myelofibrosis), leukemia (e.g. acute myeloidleukaemia and acute lymphoblastic leukemia), multiple myeloma,psoriasis, restenosis, sclerodermitis or fibrosis. In particular theterm refers to cancer, leukemia, multiple myeloma and psoriasis.

As used herein, the term ‘cancer’ refers to a malignant or benign growthof cells in skin or in body organs, for example but without limitation,breast, prostate, lung, kidney, pancreas, stomach or bowel. A cancertends to infiltrate into adjacent tissue and spread (metastases) todistant organs, for example to bone, liver, lung or the brain. As usedherein the term cancer includes both metastatic rumour cell types, suchas but not limited to, melanoma, lymphoma, leukaemia, fibrosarcoma,rhabdomyosarcoma, and mastocytoma and types of tissue carcinoma, such asbut not limited to, colorectal cancer, prostate cancer, small cell lungcancer and non-small cell lung cancer, breast cancer, pancreatic cancer,bladder cancer, renal cancer, gastric cancer, glioblastoma, primaryliver cancer, ovarian cancer, prostate cancer and uterineleiomyosarcoma.

As used herein the term ‘leukaemia’ refers to neoplastic diseases of theblood and blood forming organs. Such diseases can cause bone marrow andimmune system dysfunction, which renders the host highly susceptible toinfection and bleeding. In particular the term leukemia refers to acutemyeloid leukaemia (AML) and acute lymphoblastic leukemia (ALL).

As used herein the term ‘diseases involving impairment of cartilageturnover’ and specifically ‘diseases involving the anabolic stimulationof chondrocytes’ includes conditions such as osteoarthritis, psoriaticarthritis, juvenile rheumatoid arthritis, gouty arthritis, septic orinfectious arthritis, reactive arthritis, reflex sympathetic dystrophy,algodystrophy, Tietze syndrome or costal chondritis, fibromyalgia,osteochondritis, neurogenic or neuropathic arthritis, arthropathy,endemic forms of arthritis like osteoarthritis deformans endemica,Mseleni disease and Handigodu disease; degeneration resulting fromfibromyalgia, systemic lupus erythematosus, scleroderma and ankylosingspondylitis.

As used herein the term ‘congenital cartilage malformation(s)’ includesconditions such as hereditary chondrolysis, chondrodysplasias andpseudochondrodysplasias, in particular, but without limitation,microtia, anotia, metaphyseal chondrodysplasia, and related disorders.

As used herein the term ‘disease(s) associated with hypersecretion ofIL6’ includes conditions such as Castleman's disease, multiple myeloma,psoriasis, Kaposi's sarcoma and/or mesangial proliferativeglomerulonephritis.

‘Compound(s) of the invention’, and equivalent expressions, are meant toembrace compounds of the Formula (e) as hereinbefore described, whichexpression includes the pharmaceutically acceptable salts, and thesolvates, e.g., hydrates, and the solvates of the pharmaceuticallyacceptable salts where the context so permits. Similarly, reference tointermediates, whether or not they themselves are claimed, is meant toembrace their salts, and solvates, where the context so permits.

When ranges are referred to herein, for example but without limitation,C₁-C₈ alkyl, the citation of a range should be considered arepresentation of each member of said range.

Other derivatives of the compounds of the invention have activity inboth their acid and acid derivative forms, but in the acid sensitiveform often offers advantages of solubility, tissue compatibility, ordelayed release in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well know to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides and anhydridesderived from acidic groups pendant on the compounds of this inventionare particularly useful prodrugs. In some cases it is desirable toprepare double ester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. Particular such prodrugs are the C₁ toC₈ alkyl, C₂-C₈ alkenyl, aryl, C₇-C₁₂ substituted aryl, and C₇-C₁₂arylalkyl esters of the compounds of the invention.

As used herein, the term ‘isotopic variant’ refers to a compound thatcontains unnatural proportions of isotopes at one or more of the atomsthat constitute such compound. For example, an ‘isotopic variant’ of acompound can contain one or more non-radioactive isotopes, such as forexample, deuterium (²H or D), carbon-13 (¹³C), nitrogen-15 (¹⁵N), or thelike. It will be understood that, in a compound where such isotopicsubstitution is made, the following atoms, where present, may vary, sothat for example, any hydrogen may be ²H/D, any carbon may be ¹³C, orany nitrogen may be ¹⁵N, and that the presence and placement of suchatoms may be determined within the skill of the art. Likewise, theinvention may include the preparation of isotopic variants withradioisotopes, in the instance for example, where the resultingcompounds may be used for drug and/or substrate tissue distributionstudies. The radioactive isotopes tritium, i.e. ³H, and carbon-14, i.e.¹⁴C, are particularly useful for this purpose in view of their ease ofincorporation and ready means of detection. Further, compounds may beprepared that are substituted with positron emitting isotopes, such as¹¹C, ¹⁸F, ¹⁵O and ¹³N, and would be useful in Positron EmissionTopography (PET) studies for examining substrate receptor occupancy.

All isotopic variants of the compounds provided herein, radioactive ornot, are intended to be encompassed within the scope of the invention.

It is also to be understood that compounds that have the same molecularformula but differ in the nature or sequence of bonding of their atomsor the arrangement of their atoms in space are termed ‘isomers’. Isomersthat differ in the arrangement of their atoms in space are termedstereoisomers'.

Stereoisomers that are not mirror images of one another are termed‘diastereomers’ and those that are non-superimposable mirror images ofeach other are termed ‘enantiomers’. When a compound has an asymmetriccenter, for example, it is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center and is described by theR- and S-sequencing rules of Cahn and Prelog, or by the manner in whichthe molecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a ‘racemic mixture’.

‘Tautomers’ refer to compounds that are interchangeable forms of aparticular compound structure, and that vary in the displacement ofhydrogen atoms and electrons. Thus, two structures may be in equilibriumthrough the movement of it electrons and an atom (usually H). Forexample, enols and ketones are tautomers because they are rapidlyinterconverted by treatment with either acid or base. Another example oftautomerism is the aci- and nitro-forms of phenylnitromethane, that arelikewise formed by treatment with acid or base.

Tautomeric forms may be relevant to the attainment of the optimalchemical reactivity and biological activity of a compound of interest.

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)— or(S)— stereoisomers or as mixtures thereof.

Unless indicated otherwise, the description or naming of a particularcompound in the specification and claims is intended to include bothindividual enantiomers and mixtures, racemic or otherwise, thereof. Themethods for the determination of stereochemistry and the separation ofstereoisomers are well-known in the art.

The Compounds

The present invention is based on the discovery that inhibitors of JAKare useful for the treatment of diseases involving cartilagedegradation, bone and/or joint degradation, for example osteoarthritis;and/or conditions involving inflammation or immune responses, such asCrohn's disease, rheumatoid arthritis, psoriasis, allergic airwaysdisease (e.g. asthma, rhinitis), juvenile idiopathic arthritis, colitis,inflammatory bowel diseases, endotoxin-driven disease states (e.g.complications after bypass surgery or chronic endotoxin statescontributing to e.g. chronic cardiac failure), diseases involvingimpairment of cartilage turnover (e.g. diseases involving the anabolicstimulation of chondrocytes), congenital cartilage malformations,diseases associated with hypersecretion of IL6 and transplantationrejection (e.g. organ transplant rejection). Inhibitors of JAK can alsofind application in the treatment of proliferative diseases. Inparticular the inhibitors of JAK find application in the treatment ofcancers, especially leukaemias and solid tumours (e.g. uterineleiomyosarcoma, prostate cancer). In particular diseases involvingcartilage degradation, bone and/or joint degradation and/orinflammation, for example osteoarthritis. The present invention alsoprovides methods for the production of these compounds, pharmaceuticalcompositions comprising these compounds and methods for treatingdiseases involving cartilage degradation, bone and/or joint degradationand/or inflammation by administering a compound of the invention. Thepresent compounds may be inhibitors of one or more members of the JAKfamily; specifically they may inhibit the activity of one or more ofJAK1, JAK2, JAK3 and/or TYK2.

Accordingly, in a first aspect of the invention,1,2,4-triazolo[1,5-a]pyridine compounds are disclosed having a Formula(I):

wherein

-   -   Cy1 is selected from aryl and heteroaryl;

L1 is selected from a single bond, —O—, —C(O)—, —C[═N(R^(4a))]-,—N(R^(4a))—, —CON(R^(4a))—, —SO₂N(R^(4a))—, —S(O)₂—, —N(R^(4a))CO—, or—N(R^(4a))SO₂—;

-   -   each R¹ is independently selected from C₁-C₆ alkyl, substituted        C₁-C₆ alkyl, acyl, substituted acyl, substituted or        unsubstituted acylamino, substituted or unsubstituted C₁-C₆        alkoxy, substituted or unsubstituted amido, substituted or        unsubstituted amino, substituted sulfinyl, substituted sulfonyl,        substituted or unsubstituted aminosulfonyl, sulfonic acid,        sulfonic acid ester, carboxy, cyano, substituted or        unsubstituted C₃-C₇ cycloalkyl, substituted or unsubstituted 4-7        membered heterocycloalkyl, halo, and hydroxyl;

each R^(3a) is independently selected from C₁-C₆ alkyl, substitutedC₁-C₆ alkyl, acyl, substituted acyl, substituted or unsubstitutedacylamino, substituted or unsubstituted C₁-C₆ alkoxy, substituted orunsubstituted amido, alkoxycarbonyl, substituted alkoxycarbonyl,arylalkyloxy, substituted arylalkyloxy, substituted or unsubstitutedamino, aryl, substituted aryl, arylalkyl, substituted sulfanyl,substituted sulfinyl, substituted sulfonyl, substituted or unsubstitutedaminosulfonyl, sulfonic acid, sulfonic acid ester, azido, carboxy,cyano, substituted or unsubstituted C₃-C₇ cycloalkyl, substituted orunsubstituted 4-7 membered heterocycloalkyl, halo, substituted orunsubstituted heteroaryl, hydroxyl, nitro, and thiol;

R^(2a) is selected from substituted or unsubstituted C₁-C₆ alkyl orsubstituted or unsubstituted C₃-C₇ cycloalkyl;

R^(3b) is independently selected from substituted or unsubstituted aryl,substituted or unsubstituted C₃-C₇ cycloalkyl, substituted orunsubstituted 4-7 membered heterocycloalkyl, substituted orunsubstituted 5-10 membered heteroaryl; or R^(3b) is independentlyselected from O—R^(3c), CO—R^(3c), and CON(R^(4a))—R^(3c); and R^(3c) isindependently selected from substituted or unsubstituted aryl,substituted or unsubstituted C₃-C₇ cycloalkyl, substituted orunsubstituted 4-7 membered heterocycloalkyl, substituted orunsubstituted 5-10 membered heteroaryl;

each R^(4a), R^(4b) and R^(4c) is independently selected from H, C₁-C₆alkyl, substituted C₁-C₆ alkyl, C₃-C₇ cycloalkyl, or substituted C₃-C₇cycloalkyl;

-   -   m1 is 0, 1, or 2; m2 is 0, 1, 2, or 3; and n1 is 0, 1, 2, 3, or        4;    -   provided that        -   when L1 is —O—, —N(R^(4a))—, —CON(R^(4a))—, or            —SO₂N(R^(4a))—, and R^(3b) is other than cycloalkyl, aryl or            5-10 membered heteroaryl, then n1 is 1, 2, 3, or 4;    -   or pharmaceutically acceptable salts or solvates thereof or        solvates of the pharmaceutically acceptable salts.

In one embodiment, with respect to compounds of Formula I, m1 is 0.

In another embodiment, with respect to compounds of Formula I, R^(2a) issubstituted or unsubstituted C₃-C₇ cycloalkyl.

In a particular embodiment, with respect to compounds of Formula I,R^(2a) is cyclopropyl, cyclobutyl, or cyclopentyl.

In a further embodiment, with respect to compounds of Formula I, R^(4b)and R^(4c) are independently selected from H and Me.

In a more particular embodiment, with respect to compounds of Formula I,the compound is according to Formula II:

wherein Cy1, L1, R^(3a), R^(3b), m2, and n1 are as described previously.

In one embodiment, with respect to compounds of Formula II, Cy1 is Ph;and m2 is 0.

In a more particular embodiment, the compound is according to FormulaIII:

wherein L1, R^(3b), and n1 are as above.

In one embodiment, with respect to compounds of Formula III, R^(3b) issubstituted or unsubstituted aryl, substituted or unsubstituted 5-10membered heteroaryl, substituted or unsubstituted C₃-C₇ cycloalkyl, orsubstituted or unsubstituted 4-7 membered heterocycloalkyl.

In a particular embodiment, with respect to compounds of Formula III, L1is selected from a single bond, —O—, —N(R^(4a))—, —C(O)—,C[═N(R^(4a))]-, —CON(R^(4a))—, —SO₂N(R^(4a))—, —S(O)₂—, —N(R^(4a))SO₂—and —N(R^(4a))CO—; n1 is 0, 1, 2, 3, or 4; and R^(3b) is substituted orunsubstituted aryl, substituted or unsubstituted 5-10 memberedheteroaryl, substituted or unsubstituted C₃-C₇ cycloalkyl, substitutedor unsubstituted 4-7 membered heterocycloalkyl.

In one embodiment, with respect to compounds of Formula III, L1 isselected from — a single bond, —O—, —N(R^(4a))—, —C(O)—,—C[═N(R^(4a))]-, —CON(R^(4a))—, —SO₂N(R^(4a))—, —S(O)₂—, —N(R^(4a))SO₂—and —N(R^(4a))CO—; n1 is 0, 1, 2, 3, or 4; and R^(3b) is substituted orunsubstituted aryl or substituted or unsubstituted 5-10 memberedheteroaryl.

In a particular embodiment, with respect to compounds of Formula III, L1is selected from — a single bond, —O—, —N(R^(4a))—, —C(O)—,—C[═N(R^(4a))]-, —CON(R^(4a))—, —SO₂N(R^(4a))—, —S(O)₂—, —N(R^(4a))SO₂—and —N(R^(4a))CO—; n1 is 0, 1, 2, 3, or 4; and R^(3b) is substituted orunsubstituted phenyl, substituted or unsubstituted pyridyl, substitutedor unsubstituted pyrrolyl, substituted or unsubstituted pyrazolyl,substituted or unsubstituted imidazolyl, substituted or unsubstitutedtriazolyl, substituted or unsubstituted tetrazolyl, substituted orunsubstituted oxazolyl, substituted or unsubstituted oxadiazolyl,substituted or unsubstituted thiazolyl, substituted or unsubstitutedthiophenyl, substituted or unsubstituted indolyl, substituted orunsubstituted indazolyl, substituted or unsubstituted benzimidazolyl,substituted or unsubstituted benzofuranyl, substituted or unsubstitutedbenzodioxanyl, substituted or unsubstituted benzoxazolyl, substituted orunsubstituted quinolinyl, or substituted or unsubstituted isoquinolinyl.

In another particular embodiment, with respect to compounds of FormulaIII, L1 is selected from —O—, and —N(R^(4a))—.

In another particular embodiment, with respect to compounds of FormulaIII, L1 is —O—.

In one embodiment, the compound is according to Formula III, and-Ph-L1-(CH₂)_(n1)—R^(3b) is selected from:

wherein n2 is n1; and R^(3b), and n1 are as in Formula 1; and Cy3 is asubstituted or unsubstituted nitrogen containing 4-7-memberedheterocycloalkyl group.

In one embodiment, the compound is according to Formula III, and-Ph-L1-(CH₂)_(n1)—R^(3b) is:

wherein n2 is n1; and R^(3b), and n1 are as in Formula 1.

In one embodiment, the compound is according to Formula III, and-Ph-L1-(CH₂)_(n1)—R^(3b) is:

wherein n2 is n1; R^(3b) is independently selected from substituted orunsubstituted aryl, substituted or unsubstituted C₃-C₇ cycloalkyl,substituted or unsubstituted 4-7 membered heterocycloalkyl, substitutedor unsubstituted 5-10 membered heteroaryl; or R^(3b) is independentlyselected from O—R^(3c), CO—R^(3c), and CON(R^(4a))—R^(3c); and R^(3c) isin dependently selected from substituted or unsubstituted awl,substituted or unsubstituted C₃-C₇ cycloalkyl, substituted orunsubstituted 4-7 membered heterocycloalkyl, substituted orunsubstituted 5-10 membered heteroaryl, and n1 is 0, 1, 2, 3, or 4.

In one embodiment, the compound is according to Formula III, and-Ph-L1-(CH₂)_(n1)—R^(3b) is:

wherein n2 is n1; R^(3b) is substituted or unsubstituted 5-10 memberedheteroaryl, and n1 is 0, 1, 2, 3, or 4.

In a more particular embodiment, the compound is according to FormulaeVIa, VIb, VIc, or VId:

In a more particular embodiment, the compound is according to FormulaVIa:

-   -   or pharmaceutically acceptable salts or solvates thereof or        solvates of the pharmaceutically acceptable salts.

In one embodiment the compound is not an isotopic variant.

In one embodiment, with respect to Formula I, the compound is selectedfrom the compounds exemplified in Table 1.

In one embodiment, with respect to Formula I, the compound is compound176 from Table 1.

In certain aspects, the present invention also provides prodrugs andderivatives of the compounds according to the Formula (e) above.Prodrugs are derivatives of the compounds of the invention, which havemetabolically cleavable groups and become by solvolysis or underphysiological conditions the compounds of the invention, which arepharmaceutically active, in vivo. Such examples include, but are notlimited to, choline ester derivatives and the like, N-alkylmorpholineesters and the like.

Other derivatives of the compounds of the invention have activity inboth their acid and acid derivative forms, but the acid sensitive formoften offers advantages of solubility, tissue compatibility, or delayedrelease in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well know to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides and anhydridesderived from acidic groups pendant on the compounds of this inventionare preferred prodrugs. In some cases it is desirable to prepare doubleester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. Particularly useful are the C₁ to C₈alkyl, C)—C₈ alkenyl, aryl, C₇-C₁₂ substituted awl, and C₇-C₁₂ arylalkylesters of the compounds of the invention.

Pharmaceutical Compositions

When employed as pharmaceuticals, the compounds of the invention aretypically administered in the form of a pharmaceutical composition. Suchcompositions can be prepared in a manner well known in thepharmaceutical art and comprise at least one active compound. Generally,the compounds of this invention are administered in a pharmaceuticallyeffective amount. The amount of the compound actually administered willtypically be determined by a physician, in the light of the relevantcircumstances, including the condition to be treated, the chosen routeof administration, the actual compound-administered, the age, weight,and response of the individual patient, the severity of the patient'ssymptoms, and the like.

The pharmaceutical compositions of the invention can be administered bya variety of routes including oral, rectal, transdermal, subcutaneous,intra-articular, intravenous, intramuscular, and intranasal. Dependingon the intended route of delivery, the compounds of this invention arepreferably formulated as either injectable or oral compositions or assalves, as lotions or as patches all for transdermal administration

The compositions for oral administration can take the form of bulkliquid solutions or suspensions, or bulk powders. More commonly,however, the compositions are presented in unit dosage forms tofacilitate accurate dosing. The term “unit dosage forms” refers tophysically discrete units suitable as unitary dosages for human subjectsand other mammals, each unit containing a predetermined quantity ofactive material calculated to produce the desired therapeutic effect, inassociation with a suitable pharmaceutical excipient, vehicle orcarrier. Typical unit dosage forms include prefilled, premeasuredampules or syringes of the liquid compositions or pills, tablets,capsules or the like in the case of solid compositions. In suchcompositions, the furansulfonic acid compound is usually a minorcomponent (from about 0.1 to about 50% by weight or preferably fromabout 1 to about 40% by weight) with the remainder being variousvehicles or carriers and processing aids helpful for forming the desireddosing form.

Liquid forms suitable for oral administration may include a suitableaqueous or nonaqueous vehicle with buffers, suspending and dispensingagents, colorants, flavors and the like. Solid forms may include, forexample, any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring.

Injectable compositions are typically based upon injectable sterilesaline or phosphate-buffered saline or other injectable carriers knownin the art. As before, the active compound in such compositions istypically a minor component, often being from about 0.05 to 10% byweight with the remainder being the injectable carrier and the like.

Transdermal compositions are typically formulated as a topical ointmentor cream containing the active ingredient(s), generally in an amountranging from about 0.01 to about 20% by weight, preferably from about0.1 to about 20% by weight, preferably from about 0.1 to about 10% byweight, and more preferably from about 0.5 to about 15% by weight. Whenformulated as a ointment, the active ingredients will typically becombined with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream with,for example an oil-in-water cream base. Such transdermal formulationsare well-known in the art and generally include additional ingredientsto enhance the dermal penetration of stability of the active ingredientsor the formulation. All such known transdermal formulations andingredients are included within the scope of this invention.

The compounds of the invention can also be administered by a transdermaldevice. Accordingly, transdermal administration can be accomplishedusing a patch either of the reservoir or porous membrane type, or of asolid matrix variety.

The above-described components for orally administrable, injectable ortopically administrable compositions are merely representative. Othermaterials as well as processing techniques and the like are set forth inPart 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, MackPublishing Company, Easton, Pa., which is incorporated herein byreference.

The compounds of this invention can also be administered in sustainedrelease forms or from sustained release drug delivery systems. Adescription of representative sustained release materials can be foundin Remington's Pharmaceutical Sciences.

The following formulation examples illustrate representativepharmaceutical compositions that may be prepared in accordance with thisinvention. The present invention, however, is not limited to thefollowing pharmaceutical compositions.

Formulation 1—Tablets

A compound of the invention may be admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into240-270 mg tablets (80-90 mg of active amide compound per tablet) in atablet press.

Formulation 2—Capsules

A compound of the invention may be admixed as a dry powder with a starchdiluent in an approximate 1:1 weight ratio. The mixture may be filledinto 250 mg capsules (125 mg of active amide compound per capsule).

Formulation 3—Liquid

A compound of the invention (125 mg), may be admixed with sucrose (1.75g) and xanthan gum (4 mg) and the resultant mixture may be blended,passed through a No. 10 mesh U.S. sieve, and then mixed with apreviously made solution of microcrystalline cellulose and sodiumcarboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10mg), flavor, and color are diluted with water and added with stirring.Sufficient water may then be added with stirring. Sufficient water isthen added to produce a total volume of 5 mL.

Formulation 4—Tablets

A compound of the invention may be admixed as a dry powder with a drygelatin binder in an approximate 1:2 weight ratio. A minor amount ofmagnesium stearate is added as a lubricant. The mixture is formed into450-900 mg tablets (150-300 mg of active amide compound) in a tabletpress.

Formulation 5—Injection

A compound of the invention may be dissolved or suspended in a bufferedsterile saline injectable aqueous medium to a concentration ofapproximately 5 mg/mL.

Formulation 6—Topical

Stearyl alcohol (250 g) and a white petrolatum (250 g) are melted atabout 75° C. and then a mixture of a compound of the invention (50 g)methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate(10 g), and propylene glycol (120 g) dissolved in water (about 370 g) isadded and the resulting mixture is stirred until it congeals.

Methods of Treatment

The present compounds may be used as therapeutic agents for thetreatment of conditions in mammals that are causally related orattributable to aberrant activity of JAK. In particular, conditionsrelated to aberrant activity of one or more of JAK1, JAK2, JAK3 and/orTYK2. Accordingly, the compound of the invention and pharmaceuticalcompositions of this invention find use as therapeutics for preventingand/or treating diseases involving cartilage degradation, bone and/orjoint degradation, for example osteoarthritis; and/or conditionsinvolving inflammation or immune responses, such as Crohn's disease,rheumatoid arthritis, psoriasis, allergic airways disease (e.g. asthma,rhinitis), juvenile idiopathic arthritis, colitis, inflammatory boweldiseases, endotoxin-driven disease states (e.g. complications afterbypass surgery or chronic endotoxin states contributing to e.g. chroniccardiac failure), diseases involving impairment of cartilage turnover(e.g. diseases involving the anabolic stimulation of chondrocytes),congenital cartilage malformations, diseases associated withhypersecretion of IL6 and transplantation rejection (e.g. organtransplant rejection). Inhibitors of JAK can also find application inthe treatment of proliferative diseases. In particular the inhibitors ofJAK find application in the treatment of cancers, especially leukaemiasand solid tumours (e.g. uterine leiomyosarcoma, prostate cancer). Inparticular the conditions are selected from inflammatory conditions,conditions related to cartilage and/or joint degradation in mammalsincluding humans. In another embodiment, the compounds andpharmaceutical compositions of this invention find use as therapeuticsfor preventing and/or treating proliferative disorders in mammals,including humans. In a specific embodiment the compound of the inventionand pharmaceutical compositions thereof find use as therapeutics forpreventing and/or treating cancer in mammals including humans.

In additional method of treatment aspects, this invention providesmethods of treating a mammal susceptible to or afflicted with conditioninvolving an immune response or an autoimmune disease. The methodscomprise administering an effective condition-treating orcondition-preventing amount of one or more of the pharmaceuticalcompositions or compound of the invention herein described. In aspecific embodiment, the autoimmune disease is selected from COPD,asthma, systemic lupus erythematosis, type I diabetes mellitus andinflammatory bowel disease.

In another aspect the present invention provides the compound of theinvention for use in the treatment, prevention or prophylaxis of acondition involving an autoimmune response or an autoimmune disease. Ina specific embodiment, the autoimmune disease is selected from COPD,asthma, systemic lupus erythematosis, type I diabetes mellitus andinflammatory bowel disease.

In a method of treatment aspect, this invention provides a method oftreatment, prevention or prophylaxis in a mammal susceptible to orafflicted with diseases involving impairment of cartilage turnover (e.g.a condition associated with, or diseases involving the anabolicstimulation of chondrocytes), for example, osteoarthritis, psoriaticarthritis, juvenile rheumatoid arthritis, gouty arthritis, septic orinfectious arthritis, reactive arthritis, reflex sympathetic dystrophy,algodystrophy, Tietze syndrome or costal chondritis, fibromyalgia,osteochondritis, neurogenic or neuropathic arthritis, arthropathy,endemic forms of arthritis like osteoarthritis deformans endemica,Mseleni disease and Handigodu disease; degeneration resulting fromfibromyalgia, systemic lupus erythematosus, scleroderma and ankylosingspondylitis, which method comprises administering a therapeuticallyeffective amount of a compound according to the invention, or one ormore of the pharmaceutical compositions or compounds herein described.

In another aspect the present invention provides a compound of theinvention for use in the treatment, prevention or prophylaxis ofdiseases involving impairment of cartilage turnover (e.g. a conditionassociated with, or diseases involving the anabolic stimulation ofchondrocytes), for example, osteoarthritis, psoriatic arthritis,juvenile rheumatoid arthritis, gouty arthritis, septic or infectiousarthritis, reactive arthritis, reflex sympathetic dystrophy,algodystrophy, Tietze syndrome or costal chondritis, fibromyalgia,osteochondritis, neurogenic or neuropathic arthritis, arthropathy,endemic forms of arthritis like osteoarthritis deformans endemica,Mseleni disease and Handigodu disease; degeneration resulting fromfibromyalgia, systemic lupus erythematosus, scleroderma and ankylosingspondylitis.

The present invention also provides a method of treatment of congenitalcartilage malformations, including hereditary chondrolysis,chondrodysplasias and pseudochondrodysplasias, in particular, butwithout limitation, microtia, anotia, metaphyseal chondrodysplasia, andrelated disorders, which method comprises administering an effectiveamount of one or more of the pharmaceutical compositions or compoundsherein described.

In another aspect the present invention provides a compound of theinvention for use in the treatment, prevention or prophylaxis ofcongenital cartilage malformations, including hereditary chondrolysis,chondrodysplasias and pseudochondrodysplasias, in particular, butwithout limitation, microtia, anotia, metaphyseal chondrodysplasia, andrelated disorders.

In another aspect, this invention provides a method of treating a mammalsusceptible to or afflicted with a condition involving inflammation. Inadditional method of treatment aspects, this invention provides methodsof treating a mammal susceptible to or afflicted with diseases anddisorders which are mediated by or result in inflammation such as, forexample rheumatoid arthritis and osteoarthritis, allergic airway disease(e.g. asthma, rhinitis), juvenile idiopathic arthritis, colitis,inflammatory bowel diseases, endotoxin-driven disease states (e.g.complications after bypass surgery or chronic endotoxin statescontributing to e.g. chronic cardiac failure), and related diseasesinvolving cartilage, such as that of the joints, which method comprisesadministering an effective amount of one or more of the pharmaceuticalcompositions or compounds herein described. In a specific embodiment,the condition involving inflammation is selected from rheumatoidarthritis, osteoarthritis, allergic airway disease (e.g. asthma) andinflammatory bowel diseases. The methods comprise administering aneffective condition-treating or condition-preventing amount of one ormore of the pharmaceutical compositions or compounds herein described.

In another aspect, this invention provides a compound of the inventionfor use in the treatment, prevention or prophylaxis of a conditioninvolving inflammation. In another aspect the present invention providesa compound of the invention for use in the treatment, prevention orprophylaxis of diseases and disorders which are mediated by or result ininflammation such as, for example rheumatoid arthritis andosteoarthritis, allergic airway disease (e.g. asthma, rhinitis),juvenile idiopathic arthritis, colitis, inflammatory bowel diseases,endotoxin-driven disease states (e.g. complications after bypass surgeryor chronic endotoxin states contributing to e.g. chronic cardiacfailure), and related diseases involving cartilage, such as that of thejoints. In a specific embodiment, the condition involving inflammationis selected from rheumatoid arthritis, osteoarthritis, allergic airwaydisease (e.g. asthma) and inflammatory bowel diseases.

In further method of treatment aspects, this invention provides methodsof treating a mammal susceptible to or afflicted with a proliferativedisease, in particular cancer (e.g. solid tumors such as uterineleiomyosarcoma or prostate cancer), leukemia (e.g. AML or ALL), multiplemyeloma and/or psoriasis which methods comprise administering aneffective amount of one or more of the pharmaceutical compositions orcompounds herein described. In further method of treatment aspects, thisinvention provides methods of treating a mammal susceptible to orafflicted with cancer (e.g. solid tumors such as uterine leiomyosarcomaor prostate cancer) and/or leukemias.

In another aspect the present invention provides the compound of theinvention for use in the treatment, prevention or prophylaxis of aproliferative disease, in particular cancer (e.g. solid tumors such asuterine leiomyosarcoma or prostate cancer), leukemia (e.g. AML or ALL),multiple myeloma and/or psoriasis. In another aspect the presentinvention provides a compound of the invention for use in the treatment,prevention or prophylaxis of cancer (e.g. solid tumors such as uterineleiomyosarcoma or prostate cancer) and/or leukemias.

In further method of treatment aspects, this invention provides methodsof treating a mammal susceptible to or afflicted with diseasesassociated with hypersecretion of TL6, in particular Castleman's diseaseor mesangial proliferative glomerulonephritis which methods compriseadministering an effective amount of one or more of the pharmaceuticalcompositions or compounds herein described.

In another aspect the present invention provides a compound of theinvention for use in the treatment, prevention or prophylaxis ofdiseases associated with hypersecretion of TL6, in particularCastleman's disease or mesangial proliferative glomerulonephritis.

In further method of treatment aspects, this invention provides methodsof treating a mammal susceptible to or afflicted with transplantationrejection which methods comprise administering an effective amount ofone or more of the pharmaceutical compositions or compounds hereindescribed. In a specific embodiment, the invention provides methods oftreating organ transplant rejection.

In another aspect the present invention provides the compound of theinvention for use in the treatment, prevention or prophylaxis oftransplantation rejection. In a specific embodiment, the inventionprovides methods of treating organ transplant rejection.

As a further aspect of the invention there is provided the presentcompounds for use as a pharmaceutical especially in the treatment orprevention of the aforementioned conditions and diseases. Also providedherein is the use of the present compounds in the manufacture of amedicament for the treatment or prevention of one of the aforementionedconditions and diseases.

A particular regimen of the present method comprises the administrationto a subject in suffering from a disease involving inflammation, of aneffective amount of a compound of the invention for a period of timesufficient to reduce the level of inflammation in the patient, andpreferably terminate, the processes responsible for said inflammation. Aspecial embodiment of the method comprises administering of an effectiveamount of a compound of the invention to a subject patient sufferingfrom or susceptible to the development of rheumatoid arthritis, for aperiod of time sufficient to reduce or prevent, respectively,inflammation in the joints of said patient, and preferably terminate,the processes responsible for said inflammation.

A further particular regimen of the present method comprises theadministration to a subject in suffering from a disease conditioncharacterized by cartilage or joint degradation (e.g. osteoarthritis) ofan effective amount of a compound of the invention for a period of timesufficient to reduce and preferably terminate, the self-perpetuatingprocesses responsible for said degradation. A special embodiment of themethod comprises administering of an effective amount of a compound ofthe invention to a subject patient suffering from or susceptible to thedevelopment of osteoarthritis, for a period of time sufficient to reduceor prevent, respectively, cartilage degradation in the joints of saidpatient, and preferably terminate, the self-perpetuating processesresponsible for said degradation. In a particular embodiment saidcompounds exhibit cartilage anabolic and/or anti-catabolic properties.

Injection dose levels range from about 0.1 mg/kg/hour to at least 10mg/kg/hour, all for from about 1 to about 120 hours and especially 24 to96 hours. A preloading bolus of from about 0.1 mg/kg to about 10 mg/kgor more may also be administered to achieve adequate steady statelevels. The maximum total dose is not expected to exceed about 2 g/dayfor a 40 to 80 kg human patient.

For the prevention and/or treatment of long-term conditions, such asdegenerative conditions, the regimen for treatment usually stretchesover many months or years so oral dosing is preferred for patientconvenience and tolerance. With oral dosing, one to five and especiallytwo to four and typically three oral doses per day are representativeregimens. Using these dosing patterns, each dose provides from about0.01 to about 20 mg/kg of the compound of the invention, with particulardoses each providing from about 0.1 to about 10 mg/kg and especiallyabout 1 to about 5 mg/kg.

Transdermal doses are generally selected to provide similar or lowerblood levels than are achieved using injection doses.

When used to prevent the onset of an inflammatory condition, thecompounds of this invention will be administered to a patient at riskfor developing the condition, typically on the advice and under thesupervision of a physician, at the dosage levels described above.Patients at risk for developing a particular condition generally includethose that have a family history of the condition, or those who havebeen identified by genetic testing or screening to be particularlysusceptible to developing the condition.

The compounds of this invention can be administered as the sole activeagent or they can be administered in combination with other agents,including other compounds that demonstrate the same or a similartherapeutic activity, and that are determined to safe and efficaciousfor such combined administration. In a specific embodiment,co-administration of two (or more) agents allows for significantly lowerdoses of each to be used, thereby reducing the side effects seen.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention of adisease involving inflammation; particular agents include, but are notlimited to, immunoregulatory agents e.g. azathioprine, corticosteroids(e.g. prednisolone or dexamethasone), cyclophosphamide, cyclosporin A,tacrolimus, Mycophenolate Mofetil, muromonab-CD3 (OKT3, e.g.Orthocolone®), ATG, aspirin, acetaminophen, ibuprofen, naproxen, andpiroxicam.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention ofarthritis (e.g. rheumatoid arthritis); particular agents include but arenot limited to analgesics, non-steroidal anti-inflammatory drugs(NSAIDS), steroids, synthetic DMARDS (for example but without limitationmethotrexate, leflunomide, sulfasalazine, auranofin, sodiumaurothiomalate, penicillamine, chloroquine, hydroxychloroquine,azathioprine, and cyclosporin), and biological DMARDS (for example butwithout limitation lnfliximab, Etanercept, Adalimumab, Rituximab, andAbatacept).

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention ofproliferative disorders; particular agents include but are not limitedto: methotrexate, leukovorin, adriamycin, prenisone, bleomycin,cyclophosphamide, 5-fluorouracil, paclitaxel, docetaxel, vincristine,vinblastine, vinorelbine, doxorubicin, tamoxifen, toremifene, megestrolacetate, anastrozole, goserelin, anti-HER2 monoclonal antibody (e.g.Hcrccptin™), capecitabinc, raloxifene hydrochloride, EGFR inhibitors(e.g. Iressa (R), Tarccva™), Erbitux™), VEGF inhibitors (e.g. Avastin™),proteasome inhibitors (e.g. Velcade™), Glivec (R) or hsp90 inhibitors(e.g. 17-AAG). Additionally, a compound of the invention may beadministered in combination with other therapies including, but notlimited to, radiotherapy or surgery. In a specific embodiment theproliferative disorder is selected from cancer, myeloproliferativedisease or leukaemia.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention ofautoimmune diseases, particular agents include but are not limited to:glucocorticoids, cytostatic agents (e.g. purine analogs), alkylatingagents, (e.g. nitrogen mustards (cyclophosphamide), nitrosoureas,platinum compounds, and others), antimetabolites (e.g. methotrexate,azathioprine and mercaptopurine), cytotoxic antibiotics (e.g.dactinomycin anthracyclines, mitomycin C, bleomycin, and mithramycin),antibodies(e.g., anti-CD20, anti-CD25 or anti-CD3 (OTK3) monoclonalantibodies, Atgam® and Thymoglobuline®), cyclosporin, tacrolimus,rapamycin (sirolimus), interferons (e.g. IFN-β), TNF binding proteins(e.g. infliximab (Remicade), etanercept (Enbrel), or adalimumab(Humira)), mycophenolate, Fingolimod, Myriocin.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention oftransplantation rejection, particular agents include but are not limitedto: calcineurin inhibitors (e.g. cyclosporin or tacrolimus (FK506)),mTOR inhibitors (e.g. sirolimus, everolimus), anti-proliferatives (e.g.azathioprine, mycophenolic acid), corticosteroids (e.g. prednisolone,hydrocortisone), Antibodies (e.g. monoclonal anti-IL-2Rα receptorantibodies, basiliximab, daclizumab), polyclonal anti-T-cell antibodies(e.g. anti-thymocyte globulin (ATG), anti-lymphocyte globulin (ALG)).

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention of Asthmaand/or Rhinitis and/or COPD, particular agents include but are notlimited to: beta₂-adrenoceptor agonists (e.g. salbutamol, levalbuterol,terbutaline and bitolterol), epinephrine (inhaled or tablets),anticholinergics (e.g. ipratropium bromide), glucocorticoids (oral orinhaled) Long-acting β₂-agonists (e.g. salmeterol, formoterol,bambuterol, and sustained-release oral albuterol), combinations ofinhaled steroids and long-acting bronchodilators (e.g.fluticasone/salmeterol, budesonide/formoterol), leukotriene antagonistsand synthesis inhibitors (e.g. montelukast, zafirlukast and zileuton),inhibitors of mediator release (e.g. cromoglycate and ketotifen),biological regulators of IgE response (e.g. omalizumab), antihistamines(e.g. ceterizine, cinnarizine, fexofenadine), vasoconstrictors (e.g.oxymethazoline, xylomethazoline, nafazoline and tramazoline).

Additionally, the compound of the invention may be administered incombination with emergency therapies for asthma and/or COPD, suchtherapies include oxygen or heliox administration, nebulized salbutamolor terbutaline (optionally combined with an anticholinergic (e.g.ipratropium), systemic steroids (oral or intravenous, e.g. prednisone,prednisolone, methylprednisolone, dexamethasone, or hydrocortisone),intravenous salbutamol, nonspecific beta-agonists, injected or inhaled(e.g. epinephrine, isoetharine, isoproterenol, metaproterenol),anticholinergics (IV or nebulized, e.g. glycopyrrolate, atropine,ipratropium), methylxanthines (theophylline, aminophylline,bamiphylline), inhalation anesthetics that have a bronchodilatory effect(e.g. isoflurane, halothane, enflurane), ketamine, intravenous magnesiumsulfate.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention of IBD,particular agents include but are not limited to: glucocorticoids (e.g.prednisone, budcsonide) synthetics disease modifying, immunomodulatoryagents (e.g. methotrexate, leflunomide, sulfasalazine, mesalazine,azathioprine, 6-mercaptopurine and ciclosporin) and biological diseasemodifying, immunomodulatory agents (infliximab, adalimumab, rituximab,and abatacept).

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention of SLE,particular agents include but are not limited to: Disease-modifyingantirheumatic drugs (DMARDs) such as antimalarials (e.g. plaquenil,hydroxychloroquine), immunosuppressants (e.g. methotrexate andazathioprine), cyclophosphamide and mycophenolic acid; immunosuppressivedrugs and analgesics, such as nonsteroidal anti-inflammatory drugs,opiates (e.g. dextropropoxyphene and co-codamol), opioids (e.g.hydrocodone, oxycodone, MS Contin, or methadone) and the fentanylduragesic transdermal patch.

In one embodiment, the compound of the invention is co-administered withanother therapeutic agent for the treatment and/or prevention ofpsoriasis, particular agents include but are not limited to: topicaltreatments such as bath solutions, moisturizers, medicated creams andointments containing coal tar, dithranol (anthralin), corticosteroidslike desoximetasone (Topicort), fluocinonide, vitamin D₃ analogues (forexample, calcipotriol), Argan oiland retinoids (etretinate, acitretin,tazarotene), systemic treatments such as methotrexate, cyclosporine,retinoids, thioguanine, hydroxyurea, sulfasalazine, mycophenolatemofetil, azathioprine, tacrolimus, fumaric acid esters or biologics suchas Amevive, Enbrel, Humira, Remicade, Raptiva and ustekinumab (a IL-12and IL-23 blocker). Additionally, a compound of the invention may beadministered in combination with other therapies including, but notlimited to phototherapy, or photochemotherapy (e.g. psoralen andultraviolet A phototherapy (PUVA)).

By co-administration is included any means of delivering two or moretherapeutic-agents to the patient as part of the same treatment regime,as will be apparent to the skilled person. Whilst the two or more agentsmay be administered simultaneously in a single formulation this is notessential. The agents may be administered in different formulations andat different times.

General Synthetic Procedures General

The compounds of the invention can be prepared from readily availablestarting materials using the following general methods and procedures.It will be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvent used, butsuch conditions can be determined by one skilled in the art by routineoptimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary to prevent certainfunctional groups from undergoing undesired reactions. The choice of asuitable protecting group for a particular functional group as well assuitable conditions for protection and deprotection are well known inthe art. For example, numerous protecting groups, and their introductionand removal, are described in T. W. Greene and P. G. M. Wuts, ProtectingGroups in Organic Synthesis, Second Edition, Wiley, New York, 1991, andreferences cited therein.

The following methods are presented with details as to the preparationof representative bicycloheteroaryls that have been listed hereinabove.The compounds of the invention may be prepared from known orcommercially available starting materials and reagents by one skilled inthe art of organic synthesis.

All reagents were of commercial grade and were used as received withoutfurther purification, unless otherwise stated. Commercially availableanhydrous solvents were used for reactions conducted under inertatmosphere. Reagent grade solvents were used in all other cases, unlessotherwise specified. Column chromatography was performed on silica gel60 (35-70 μm). Thin layer chromatography was carried out usingpre-coated silica gel F-254 plates (thickness 0.25 mm) ¹H NMR spectrawere recorded on a Bruker DPX 400 NMR spectrometer (400 MHz). Chemicalshifts (δ) for 1H NMR spectra are reported in parts per million (ppm)relative to tetramethylsilane (δ0.00) or the appropriate residualsolvent peak, i.e. CHCl3 (δ7.27), as internal reference. Multiplicitiesare given as singlet (s), doublet (d), triplet (t), quartet (q),multiplet (m) and broad (br). Coupling constants (J) are given in Hz.Electrospray MS spectra were obtained on a Micromass platform LC/MSspectrometer. Column Used for all LCMS analysis: Waters Acquity HPLC BEHC18 1.7 μm, 2.1 mm ID×50 mm L (Part No. 186002350)). PreparativeHPLC:Waters XBridge Prep C18 5 μm ODB 19 mm ID×100 mm L (Part No.186002978). All the methods are using MeCN/H₂O gradients. H₂O containseither 0.1% TFA or 0.1% NH₃.

List of abbreviations used in the experimental section:

DCM Dichloromethane

DiPEA N,N-diisopropylethylamine

MeCN Acetonitrile

BOC tert-Butyloxy-carbonyl

DMF N,N-dimethylformamide

TFA Trifluoroacetic acid

THF Tetrahydrofuran

NMR Nuclear Magnetic Resonance

DMSO Dimethylsulfoxide

DPPA Diphenylphosphorylazide

LC-MS Liquid Chromatography-Mass Spectrometry

Ppm part-per-million

EtOAc ethyl acetate

APCI atmospheric pressure chemical ionization

Rt retention time

singlet

br s broad singlet

m multiplet

d doublet

PdCl₂dppf [1,1′-Bis(diphenylphosphino)ferrocene] dichloropalladium(II)

TEA Triethylamine

Synthetic Preparation of Compounds of the Invention

A compound of the invention can be produced according to the followingscheme.

General Synthetic Method

General 1.1.1 1-(6-Bromo-pyridin-2-yl)-3-carboethoxy-thiourea (2)

To a solution of 2-amino-6-bromopyridine (1) (253.8 g, 1.467 mol) in DCM(2.5 L) cooled to 5° C. is added ethoxycarbonyl isothiocyanate (173.0mL, 1.467 mol) dropwise over 15 min. The reaction mixture is thenallowed to warm to room temp. (20° C.) and stirred for 16 h. Evaporationin vacuo gives a solid which may be collected by filtration, thoroughlywashed with petrol (3×600 mL) and air-dried to afford (2). The thioureamay be used as such for the next step without any purification. ¹H (400MHz, CDCl₃) δ 12.03 (1H, br s, NH), 8.81 (1H, d, J 7.8 Hz, H-3), 8.15(1H, br s, NH), 7.60 (1H, t, J 8.0 Hz, H-4), 7.32 (1H, dd, J 7.7 and 0.6Hz, H-5), 4.31 (2H, q, J 7.1 Hz, CH₂), 1.35 (3H, t, J 7.1 Hz, CH₃).

1.1.2 5-Bromo-[1,2,4]triazolo[1,5-a]pyridin-2-ylamine (3)

To a suspension of hydroxylamine hydrochloride (101.8 g, 1.465 mol) inEtOH/MeOH (1:1, 900 mL) is added N,N-diisopropylethylamine (145.3 mL,0.879 mol) and the mixture is stirred at room temp. (20° C.) for 1 h.1-(6-Bromo-pyridin-2-yl)-3-carboethoxy-thiourea (2) (89.0 g, 0.293 mol)may then be added and the mixture slowly heated to reflux (Note: bleachscrubber is required to quench H₂S evolved). After 3 h at reflux, themixture is allowed to cool and filtered to collect the precipitatedsolid. Further product may be collected by evaporation in vacuo of thefiltrate, addition of H₂O (250 mL) and filtration. The combined solidsare washed successively with H₂O (250 mL), EtOH/MeOH (1:1, 250 mL) andEt₂O (250 mL) then dried in vacuo to afford the triazolopyridinederivative (3) as a solid. The compound may be used as such for the nextstep without any purification. ¹H (400 MHz, DMSO-d₆) δ 7.43-7.34 (2H, m,2×aromatic-H), 7.24 (1H, dd, J 6.8 and 1.8 Hz, aromatic-H), 6.30 (2H,br, NH₂); m/z 213/215 (1:1, M+H¹, 100%).

1.1.3 General Procedure for Mono-Acylation to Afford Intermediate (4)

To a solution of the 2-amino-triazolopyridine (3) (7.10 g, 33.3 mmol) indry CH₃CN (150 mL) at 5° C. is added Et₃N (11.6 mL, 83.3 mmol) followedby the appropriate acid chloride (83.3 mmol). The reaction mixture isthen allowed to warm to ambient temperature and stirred until allstarting material (3) is consumed. If required, further Et₃N (4.64 mL,33.3 mmol) and the acid chloride (33.3 mmol) may be added to ensurecomplete reaction. Following solvent evaporation in vacuo the resultantresidue is treated with 7 N methanolic ammonia solution (50 mL) andstirred at ambient temp. (for 1-16 h) to hydrolyse any bis-acylatedproduct. Product isolation is made by removal of volatiles in vacuofollowed by trituration with Et₂O (50 mL). The solids may be collectedby filtration, washed with H₂O (2×50 mL), acetone (50 mL) and Et₂O (50mL), then dried in vacuo to give the required acyl intermediate (4). Insome cases column chromatography (petrol/EtOAc) may be required toobtain pure compounds.

Method A 1.1.4 Preparation of Compounds of the Invention Via SuzukiCoupling (5)

An appropriate boronic acid (2 eq.) is added to a solution of bromointermediate (4) in 1,4-dioxane/water (5:1). K₂CO₃ (2 eq.) and PdCl₂dppf(5%) are added to the solution. The resulting mixture is then heated ina microwave at 140° C. for 30 min (This reaction can also be carried outby traditional heating in an oil bath at 90° C. for 16 h under N2).Water is added and the solution is extracted with ethyl acetate. Theorganic layers are dried over MgSO₄ and evaporated in vacuo. The finalcompound is obtained after purification by flash chromatography.

Method C

wherein R^(4a), R^(4b), R^(4c), R^(3b), and n1 are as described herein.

Reaction of Alkylation (General Method)

Cyclopropanecarboxylic acid[5-(4-hydroxy-phenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl]-amide (1.1 eq)obtained by Method A and K₂CO₃ (5 eq) (or AgCO₃) are dissolved in DMFunder N₂ and the appropriate alkylating agent (1.1 eq) is addeddropwise. The resulting suspension is heated at 50° C. for 16 h. Afterthis time, the reaction is complete. The compound is extracted withEtOAc and water, washed with brine and dried over MgSO₄. Organic layersare filtered and evaporated. The final compound is isolated bypreparative HPLC. Preparative HPLC: Waters XBridge Prep C18 5 μm ODB 19mm ID×100 mm L (Part No. 186002978). All the methods are using MeCN/H₂Ogradients. H₂O contains either 0.1% TFA or 0.1% NH₃.

Method E

wherein R^(4a),R^(4b), R^(4c),R^(3b) and n1 are as described herein.

Reductive Amination (General Method)

The appropriate aldehyde (2 eq.), the aniline derivative (1 eq.)obtained by Method A and Ti(OPr)₄ are mixed and stirred at roomtemperature for 3 hrs. The mixture is diluted in ethanol and Na(CN)BH₃(1 eq.) was added. The resulting solution is stirred at room temperaturefor 16 hrs. The mixture is diluted in water and filtered. The filtrateis washed with ethanol. The combined solvent phases are evaporated undervacuum. The final compound is isolated by preparative HPLC.

Preparative HPLC: Waters XBridge Prep C18 5 μm ODB 19 mm ID×100 mm L(Part No. 186002978). All the methods are using MeCN/H₂O gradients. H₂Ocontains either 0.1% TFA or 0.1% NH₃.

Method F

wherein Ar is Cy1-L1-(CR^(4b)R^(4c))_(n1)—R^(3b); and Cy1, L1, n1,R^(3b), R^(4b), and R^(4c) are as described herein.

N-(5-Bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-acetamide

To a solution of the 5-bromo-2-amino-triazolopyridine (1 eq.) in dryCH₃CN at 5° C. is added Et₃N (2.5 eq.) followed by acetyl chloride (2.5eq.). The reaction mixture is then allowed to warm to ambienttemperature and stirred until all starting material is consumed. Ifrequired, further Et₃N (1 eq.) and acid chloride (1 eq.) are added toensure complete reaction. Following solvent evaporation in vacuo theresultant residue is treated with 7 N methanolic ammonia solution andstirred at ambient temp. (for 16 h) to hydrolyse any bis-acylatedproduct. Product isolation is made by removal of volatiles in vacuofollowed by addition of water and extraction with ethyl acetate. Theorganic phase is then dried over MgSO₄, evaporated in vacuo. Thecompound is used without further purification.

Suzuki Reaction (General Method)

Boronic acids (2 eq.) is added to a solution ofN-(5-Bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-acetamide in1,4-dioxane/water (5:1). K₂CO₃ (2 eq.) and Pd(dppf)Cl₂ (5%)(dppf=1,1′-Bis(diphenylphosphino)ferrocene) are added to the solution.The resulting mixture is then heated in a microwave oven (CEM discover)in a sealed tube at 140° C. for 30 min. Water is added and the solutionis extracted with ethyl acetate. The organic layers are dried over MgSO₄and evaporated in vacuo. The final compound is obtained afterpurification by preparative HPLC. Analytical: Waters Acquity HPLC BEHC18 1.7 μm, 2.1 mm ID×50 mm L (Part No. 186002350).

Preparative HPLC: Waters XBridge Prep C18 5 μm ODB 19 mm ID×100 mm L(Part No. 186002978). All the methods are using MeCN/H₂O gradients. H₂Ocontains either 0.1% TFA or 0.1% NH₃.

Method L

L.1 Nucleophilic Aromatic Substitution (General Method)

Cyclopropanecarboxylic acid{5-[4-(6-chloro-pyridin-3-ylmethoxy)-phenyl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl}-amideprepared by method C (1 eq), an appropriate amine, (1.5 eq.) are mixedin DMSO in a sealed tube. The reaction is heated at 100° C. for 24hours. Once all the SM disappeared by LCMS, water is added to thereaction mixture and the organics is extracted with ethyl acetate. Theorganic layer is dried over MgSO₄ and evaporator under vacuum. The finalcompound is isolated by preparative HPLC. Analytical: Waters AcquityHPLC BEH C18 1.7 μm, 2.1 mm ID×50 mm L (Part No. 186002350)

Preparative HPLC: Waters XBridge Prep C18 5 μm ODB 19 mm ID×100 mm L(Part No. 186002978). All the methods are using MeCN/H₂O gradients. H₂Ocontains either 0.1% TFA or 0.1% NH₃.

Method N

N.1 Cyclopropanecarboxylic acid{514-(nitril-arylmethoxy)-phenyl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl}-amide

Pd(PPh₃)₄ (0.04 mmol) is added to degassed solution ofcyclopropanecarboxylic acid{5-[4-(haloaryl-3-ylmethoxy)-phenyl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl}-amideprepared by method B (0.24 mmol) and Zn(CN)₂ (0.24 mmol) in DMF (1 mL).The reaction mixture is exposed to microwave radiation (W:150 W; T: 150°C.) for 30 min. The mixture is diluted with ethyl acetate and washedwith water. The organic layer is dried over MgSO₄, filtered and thesolvent is removed under vacuum. The compound is purified by preparativeHPLC to afford the product (30% to 50%).

Synthesis of Representative Compounds of the Invention Compound 57:Cyclopropanecarboxylic acid[5-(6-benzyloxy-pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl]-amide

At 0° C. and under N₂ atmosphere, Benzyl alcohol (2 eq) in a solution ofTHF was treated with NaH 60% in mineral oil (4 eq) for 30 min. Thencyclopropanecarboxylic acid[5-(6-chloro-pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl]-amideprepared by method A was added to the solution and the mixture wasstirred at 70° C. for 3 hours. The reaction was completed. The reactionmixture was quenched with water and the compound was extracted withEtOAc. The compound was washed with brine, dried on MgSO₄, filtrated andconcentrated. Compound was purified on Prep HPLC.

Compound 176:N-(5-(4-((6-cyanopyridin-3-yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin-2-yl)cyclopropanecarboxamide176.1: Synthesis of5-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenoxymethyl]-pyridine-2-carbonitrile

To 4-hydroxyphenylboronic acid pinacol ester (25 g; 0.11 mol; 1.0equiv.) in acetone (250 mL) at room temperature were added under argon5-chloromethyl-pyridine-2-carbonitrile (19 g; 0.12 mol; 1.1 equiv.) andcesium carbonate (73.9 g, 0.22 mol; 2 equiv.). The reaction mixture washeated for 4 hours at reflux. The mixture was then cooled to roomtemperature, the acetone was evaporated. Water (200 mL) was added andthe product was extracted with EtOAc (3×200 mL). The organic layer wasdried over magnesium sulfate, filtered and concentrated to dryness. Theresulting residue was purified by chromatography over silica gel(petrol:EtOAc 10:1) to afford the expected boronate as a white solid.

176.2: Synthesis of Cyclopropanecarboxylic acid{5-[4-(6-cyano-pyridin-3-ylmethoxy)-phenyl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl}-amide

5-[4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenoxymethyl]-pyridine-2-carbonitrile(10 g, 0.03 mol, 1.1 equiv.) was added to a solution ofcyclopropanecarboxylic acid(5-bromo-[1,2,4]triazolo[1,5-a]pyridin-2-yl)-amide (7.6 g, 0.027 mol) in1,4-dioxane/water (4:1; 70 mL). K₂CO₃ (7.45, 0.054 mol, 2 eq.) andPdCl₂dppf (5%) were added to the solution. The resulting mixture wasthen heated in an oil bath at 90° C. under N₂ until completion(monitored by LCMS). 1,4-Dioxane was removed under vacuum, andwater/EtOAc were added and the solid was filtered. The obtained solidwas dissolved in methanol/DCM, dried over MgSO4 and the final compoundwas obtained after purification by flash chromatography, eluted withneat EtOAc as a white solid

Compound 192:5-{4-[2-(Cyclopropanecarbonyl-amino)-[1,2,4]triazolo[1,5-a]pyridin-5-yl]-phenoxymethyl}-pyridine-2-carboxylicacid amide

To a solution of cyclopropanecarboxylic acid{5-[4-(6-cyano-pyridin-3-ylmethoxy)-phenyl]-[1,2,4]triazolo[1,5-a]pyridin-2-yl}-amide(30 mg, 0.073 mmol, 1 equiv) and K₂CO₃ (10 mg, 0.073 mmol, 1 equiv) inDMSO (0.2 mL) at 10° C., 30% H₂O₂ (17 μL, 0.146 mmol, 2 equiv) was addeddropwise. After stirring at room temperature the mixture for 4 h, it wasdiluted with DMSO and filtered. The filtrate was submitted forpreparative HPLC purification: HPLC system (XBridge™ Prep C18, 5 μm,19×100 mm column); 8 min LC; flow: 20 mL/min; gradient: from 30% to 70%acetonitrile in water 0.1% TFA; isolating the final pure product.

The exemplary compounds that have been or can be prepared according tothe synthetic methods described herein are listed in Table I below. TheNMR spectral data of some representative compounds of the invention isgiven in Table II.

TABLE I Cpd MS # Structure Name Method MW Mes'd  12

N-(5-(4-(benzyloxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide A 384.44 385.10  14

N-(5-(3-(benzyloxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide A 384.44 385.20  15

N-(5-(4-(benzyloxy)-3- fluorophenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide A 402.43 403.10  16

N-(5-(2-(benzyloxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide A 384.44 385.20  36

N-(5-(4-(pyridin-3- ylmethoxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide C 385.43 408.0  (M⁺ + Na)  37

N-(5-(4-(pyridin-2- ylmethoxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide C 385.43 408.0  (M⁺ + Na)  38

N-(5-(4-(3- (trifluoromethoxy)benzyloxy) phenyl)-[1,2,4]triazolo[1,5-a]pyridin-2- yl)cyclopropanecarboxamide C 468.44 469.00  46

N-(5-(4-phenoxyphenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide A 370.41 371.00  57

N-(5-(6-(benzyloxy)pyridin- 3-yl)-[1,2,4]triazolo[1,5- a]pyridin-2-yl)cyclopropanecarboxamide Described above 385.43 386.00  72

N-(5-(4- (benzylamino)phenyl)- [1,2,4]triazolo[1,5-a]pyridin- 2-yl)cyclopropanecarboxamide E 383.46 384.00  78

N-(5-(4-((6- (trifluoromethyl)pyridin-3- yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide C 453.43454.00  92

N-(5-(4-(4- acetamidobenzyloxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin-2- yl)cyclopropanecarboxamide C 441.49 442.00 127

N-(5-(4-(2-(pyridin-3- yl)ethoxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin-2- yl)cyclopropanecarboxamide C 399.45 400.0  163

N-(5-(4-(benzyloxy)phenyl)- [1,2,4]triazolo[1,5-a]pyridin-2-yl)acetamide F 358.40 359.0  165

N-(5-(4-((6- morpholinopyridin-3- yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide L 470.53471.1  167

N-(5-(4-((6-(4- methylpiperazin-1- yl)pyridin-3- yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide L 483.58484.1  174

N-(5-(4-((6-(pyrrolidin-1- yl)pyridin-3- yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide L 454.53455.1  176

N-(5-(4-((6-cyanopyridin-3- yl)methoxy)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide Methoddescribed above 410.44 411.0  182

N-(5-(4-(pyridin-3- ylmethylamino)phenyl)-[1,2,4]triazolo[1,5-a]pyridin- 2- yl)cyclopropanecarboxamide E 384.44385.0  190

methyl 6-((4-(2- (cyclopropanecarboxamido)-[1,2,4]triazolo[1,5-a]pyridin- 5-yl) phenylcyclopropanecarboxamido)-[1,2,4]triazolo[1,5- a]pyridin-5- yl)phenoxy)methyl)nicotinate C 443.47444.0  192

5-((4-(2- (cyclopropanecarboxamido)- [1,2,4]triazolo[1,5-a]pyridin-5-yl) phenylcyclopropanecarboxamido)- [1,2,4]triazolo[1,5- a]pyridin-5-yl)phenoxy)methyl)picolinamide Described above 428.45 429.1  197

Cyclopropanecarboxylic acid {5-[4-(6-methyl-pyridin-3-ylmethoxy)-phenyl]- [1,2,4]triazolo[1,5-a]pyridin- 2-yl}-amide C 399.45400.1  198

Cyclopropanecarboxylic acid {5-[4-(6-chloro-pyridin-3-ylmethoxy)-phenyl]- [1,2,4]triazolo[1,5-a]pyridin- 2-yl}-amide C 419.87420.0 

TABLE II NMR Data of Representative Compounds of the Invention Cpd # (δ)NMR data 12 (¹H, CDCl₃) 8.70 (1H, b, NH), 7.97 (2H, d, ArH), 7.60-7.30(7H, m, ArH), 7.15 (2H, m, ArH), 7.07 (1H, m, ArH), 5.17 (2H, s, CH₂),1.60 (1H, under water peak, CH), 1.21 (2H, m, CH₂), 0.94 (2H, m, CH₂) 15(¹H, DMSO) 11.04 (1H, br, NH), 8.08 (1H, d, ArH), 7.88 (1H, d, ArH),7.68 (2H, m, ArH), 7.51 (2H, d, ArH), 2.48-7.30 (5H, m, ArH), 5.32 (2H,s, CH₂), 2.02 (1H, br, CH), 0.83 (4H, m, 2 × CH₂) 36 (¹H, DMSO) 11.03(1H, br, NH), 8.62 (1H, m, ArH), 8.02 (2H, d, ArH), 7.89 (1H, m, ArH),7.66 (2H, m, ArH), 7.58 (1H, d, ArH), 7.40 (1H, dd, ArH), 7.27 (1H, d,ArH), 7.21 (2H, d, ArH), 5.31 (2H, s, CH₂), 2.02 (1H, br, CH), 0.83 (4H,m, 2 × CH₂) 37 (¹H, DMSO) 11.01 (1H, br, NH), 8.04 (2H, d, ArH), 7;68(2H, m, ArH), 7.55 (2H, m, ArH), 7.50 (1H, s, ArH), 7.38 (1H, d, ArH),7.28 (1H, d, ArH), 7.20 (2H, d, ArH), 5.30 (2H, s, CH₂), 2.02 (1H, br,CH), 0.82 (4H, m, 2 × CH₂) 38 (¹H, DMSO) 11.01 (1H, br, NH), 8.00 (2H,d, ArH), 7.69 (2H, d, ArH), 7.69 (1H, dd, ArH), 7.64 (1H, d, ArH), 7.26(1H, d, ArH), 7.11 (2H, d, ArH), 4.07 (2H, d, CH₂), 2.78 (1H, m, CH),2.2-1.8 (7H, m, CH, 3 × CH₂), 0.83 (4H, m, 2 × CH₂) 57 (¹H, DMSO) 11.18(1H, br, NH), 8.01 (2H, d, ArH), 7.68 (2H, m, ArH), 7.26 (1H, d, ArH),7.09 (2H, d, ArH), 4.12 (2H, t, CH₂), 2.86 (2H, t, CH₂), 2.28 (6H, s, 2× CH₃), 2.02 (1H, br, CH), 0.81 (4H, m, 2 × CH₂). 72 (¹H, DMSO) 10.95(1H, br, NH), 7.85 (2H, d, ArH), 7.61 (1H, dd, ArH), 7.51 (1H, d, ArH),7.40 (4H, m, ArH), 7.36 (1H, m, ArH), 7.24 (1H, d, ArH), 7.09 (4H, m,ArH), 6.88 (1H, m, ArH), 6.70 (2H, d, ArH), 4.37 (2H, d, CH₂), 2.02 (1H,br, CH), 0.81 (4H, m, 2 × CH₂) 73 (¹H, DMSO) 11.05 (1H, s, NH), 8.09(2H, d, ArH), 8.02 (2H, d, ArH), 7.3-7.56 (7H, m, ArH), 7.36 (1H, m,ArH), 4.54 (1H, b, CH), 3.80 (2H, m, CH₂), 3.06 (1H, br, CH), 2.02-1.80(4H, br, 2 × CH, CH₂), 1.58 (2H, m, CH₂), 0.81 (4H, m, 2 × CH₂) 78 (¹H,DMSO) 11.05 (1H, s, NH), 8.09 (2H, d, ArH), 8.02 (2H, d, ArH), 7.3-7.56(7H, m, ArH), 7.36 (1H, m, ArH), 4.54 (1H, b, CH), 3.80 (2H, m, CH₂),3.06 (1H, br, CH), 2.02-1.80 (4H, br, 2 × CH, CH₂), 1.58 (2H, m, CH₂),0.81 (4H, m, 2 × CH₂) 92 (¹H, DMSO) 11.03 (1H, br, NH), 10.01 (1H, br,NH), 8.00 (2H, d, ArH), 7.65 (4H, m, ArH), 7.41 (2H, d, ArH), 7.27 (1H,m, ArH), 7.17 (2H, d, ArH), 5.15 (2H, s, CH₂), 2.04 (4H, m, CH₃, CH),0.82 (4H, m, 2 × CH₂). 127 (¹H, DMSO), 10.98 (1H, s, NH), 8.63 (1H, s,ArH), 8.51 (1H, m, ArH), 8.00 (2h, d, ArH), 7.91 (1H, d, ArH), 7.64 (2H,m, ArH), 7.46 (1H, m, ArH), 7.24 (1H, m, ArH), 7.13 (2H, m, ArH), 4.36(2H, t, CH₂), 3.14 (2H, t, CH₂), 2.04 (1H, br, CH), 0.82 (4H, m, 2 ×CH₂). 163 (¹H, DMSO) 10.72 (1H, br, NH), 7.99 (2H, m, ArH), 7.65 (2H, b,ArH), 7.48 (4H, b, ArH), 7.18 (2H, m, ArH), 5.21 (2H, d, CH₂), 2.12 (3H,br, CH₃) 165 (¹H, DMSO) 8.26 (1H, d, ArH), 8.01 (2H, d, ArH), 7.68 (2H,m, ArH), 7.62 (1H, d, ArH), 7.26 (1H, d, ArH), 7.17 (2H, d, ArH), 6.87(1H, d, ArH), 5.10 (2H, s, CH₂), 3.69 (4H, t, 2 × CH₂), 3.46 (4H, t, 2 ×CH₂), 2.07 (1H, br, CH), 0.81 (4H, m, 2 × CH₂) 167 (¹H, DMSO) 11.00 (1H,br, NH), 10.03 (1H, br, NH), 8.31 (1H, d, ArH), 8.03 (2H, d, ArH), 7.76(1H, d, ArH), 7.69 (1H, m, ArH), 7.63 (1H, d, ArH), 7.26 (1H, m, ArH),7.18 (2H, m, ArH), 7.00 (1H, d, ArH), 5.12 (2H, s, CH₂), 4.41 (2H, br,CH₂), 3.49 (2H, br, CH₂), 3.15 (4H, br, 2 × CH₂), 2.84 (3H, s, CH₃),2.04 (1H, br, CH), 0.82 (4H, m, 2 × CH₂) 174 (¹H, DMSO) 11.02 (1H, br,NH), 8.14 (1H, s, ArH), 8.03 (3H, m, ArH), 7.66 (2H, m, ArH), 7.27 (1H,d, ArH), 7.20 (2H, dd, ArH), 7.10 (1H, d, ArH), 5.17 (2H, s, CH₂), 3.53(4H, t, 2 × CH₂), 2.02 (5H, t, 2 × CH₂, CH), 0.82 (4H, m, 2 × CH₂). 176(¹H, DMSO) 11.01 (1H, br, NH), 8.89 (1H, s, ArH), 8.16 (1H, d, ArH),8.09 (1H, d, ArH), 8.04 (2H, d, ArH), 7.67 (2H, m, ArH), 7.27 (1H, d,ArH), 7.23 (2H, d, ArH), 5.41 (2H, s, CH₂), 2.04 (1H, br, CH), 0.81 (4H,m, 2 × CH₂). 182 (¹H, DMSO) 10.94 (1H, b, NH), 8.62 (1H, s, ArH), 8.46(1H, d, ArH), 7.86 (2H, d, ArH), 7.78 (1H, d, ArH), 7.62 (1H, dd, ArH),7.51 (1H, d, ArH), 7.37 (1H, m, ArH), 6.89 (1H, m, NH), 6.73 (2H, d,ArH), 6.57 (1H, s, ArH), 4.42 (2H, d, CH₂), 2.05 (1H, br, CH), 0.81 (4H,m, 2 × CH₂). 190 (¹H, DMSO) 11.04 (1H, br, NH), 9.11 (1H, s, ArH), 8.38(1H, d, ArH), 8.02 (2H, d, ArH), 7.69 (3H, m, ArH), 7.22 (3H, m, ArH),5.41 (2H, s, ArH), 3.90 (3H, s, ArH), 2.04 (1H, br, CH), 0.81 (4H, m, 2× CH₂). 192 (¹H, DMSO) 11.07 (1H, b, NH), 8.76 (1H, s, ArH), 8.16 (1H,br, ArH), 8.09 (2H, s, ArH), 8.03 (2H, d, ArH), 7.68 (3H, m, ArH), 7.28(1H, d, ArH), 7.23 (2H, d, ArH), 5.38 (2H, s, CH₂), 2.02 (1H, br, CH),0.81 (4H, m, 2 × CH₂). 197 (1H, DMSO) 10.99 (1H, b, NH), 8.57 (1H, d,ArH), 8.02 (2H, d, ArH), 7.79 (1H, dd, ArH), 7.69 (1H, dd, ArH), 7.63(1H, dd, ArH), 7.29 (1H, d, ArH), 7.26 (1H, dd, ArH), 7.20 (2H, d, ArH),5.22 (2H, s, CH₂), 2.50 (3H, s, CH₃), 2.03 (1H, b, CH), 0.81 (4H, m,CH₂). 198 (1H, DMSO) 10.99 (1H, b, NH), 8.56 (1H, m, ArH), 8.03 (2H, d,ArH), 7.99 (1H, dd, ArH), 7.69 (1H, dd, ArH), 7.63 (1H, dd, ArH), 7.58(1H, d, ArH), 7.27 (1H, dd, ArH), 7.21 (2H, d, ArH), 5.29 (2H, s, CH₂),2.04 (1H, b, CH), 0.81 (4H, m, CH₂).

BIOLOGICAL EXAMPLES Example 1 In Vitro Assays Example 1.1 JAK1Inhibition Assay

Recombinant human JAK1 catalytic domain (amino acids 850-1154; catalognumber 08-144) was purchased from Carna Biosciences. 10 ng of JAK1 wasincubated with 12.5 μg polyGT substrate (Sigma catalog number P0275) inkinase reaction buffer (15 mM Tris-HCl pH 7.5, 1 mM DTT, 0.01% Tween-20,10 mM MgCl₂, 2 μM non-radioactive ATP, 0.25 μCi 33P-gamma-ATP (GEHealthcare, catalog number AH9968) final concentrations) with or without5 μL containing test compound or vehicle (DMSO, 1% final concentration),in a total volume of 25 μL, in a polypropylene 96-well plate (Greiner,V-bottom). After 45 min at 30° C., reactions were stopped by adding of25 μL/well of 150 mM phosphoric acid. All of the terminated kinasereaction was transferred to prewashed (75 mM phosphoric acid) 96 wellfilter plates (Perkin Elmer catalog number 6005177) using a cellharvester (Perkin Elmer). Plates were washed 6 times with 300 μL perwell of a 75 mM phosphoric acid solution and the bottom of the plateswas sealed. 40 μL/well of Microscint-20 was added, the top of the plateswas sealed and readout was performed using the Topcount (Perkin Elmer).Kinase activity was calculated by subtracting counts per minute (cpm)obtained in the presence of a positive control inhibitor (10 μMstaurosporine) from cpm obtained in the presence of vehicle. The abilityof a test compound to inhibit this activity was determined as:

Percentage inhibition=((cpm determined for sample with test compoundpresent−cpm determined for sample with positive controlinhibitor)divided by(cpm determined in the presence of vehicle−cpmdetermined for sample with positive control inhibitor))*100%.

Dose dilution series were prepared for the compounds enabling thetesting of dose-response effects in the JAK1 assay and the calculationof the IC₅₀ for each compound. Each compound was routinely tested atconcentration of 20 μM followed by a ⅓ serial dilution, 8 points (20μM-6.67 μM-2.22 μM-740 nM-247 nM-82 nM-27 nM-9 nM) in a finalconcentration of 1% DMSO. When potency of compound series increased,more dilutions were prepared and/or the top concentration were lowered(e.g. 5 μM, 1 μM).

Semi-Quantitative Score:

-   -   *>501 nM    -   **101-500 nM    -   ***0.1-100 nM

TABLE III JAK1 IC₅₀ Values of Compounds Cpd # JAK1 12 *** 14 ** 15 ***36 *** 37 *** 38 ** 57 *** 72 *** 78 *** 92 *** 127 ** 163 *** 165 **167 ** 174 * 176 *** 182 *** 190 *** 192 *** 197 *** 198 ***

Example 1.2 JAK2 Inhibition Assay

Recombinant human JAK2 catalytic domain (amino acids 808-1132; catalognumber PV4210) was purchased from Invitrogen. 0.025 mU of JAK2 wasincubated with 2.5 μg polyGT substrate (Sigma catalog number P0275) inkinase reaction buffer (5 mM MOPS pH 7.5, 9 mM MgAc, 0.3 mM EDTA, 0.06%Brij and 0.6 mM DTT, 1 μM non-radioactive ATP, 0.25 μCi 33P-gamma-ATP(GE Healthcare, catalog number AH9968) final concentrations) with orwithout 5 μL containing test compound or vehicle (DMSO, 1% finalconcentration), in a total volume of 25 μL, in a polypropylene 96-wellplate (Greiner, V-bottom). After 90 min at 30° C., reactions werestopped by adding of 25 μL/well of 150 mM phosphoric acid. All of theterminated kinase reaction was transferred to prewashed (75 mMphosphoric acid) 96 well filter plates (Perkin Elmer catalog number6005177) using a cell harvester (Perkin Elmer). Plates were washed 6times with 300 mL per well of a 75 mM phosphoric acid solution and thebottom of the plates was sealed. 40 μL/well of Microscint-20 was added,the top of the plates was sealed and readout was performed using theTopcount (Perkin Elmer). Kinase activity was calculated by subtractingcounts per minute (cpm) obtained in the presence of a positive controlinhibitor (10 μM staurosporine) from cpm obtained in the presence ofvehicle. The ability of a test compound to inhibit this activity wasdetermined as:

Percentage inhibition=((cpm determined for sample with test compoundpresent−cpm determined for sample with positive controlinhibitor)divided by(cpm determined in the presence of vehicle−cpmdetermined for sample with positive control inhibitor))*100%.

Dose dilution series were prepared for the compounds enabling thetesting of dose-response effects in the JAK2 assay and the calculationof the IC₅₀ for each compound. Each compound was routinely tested atconcentration of 20 μM followed by a ⅓ serial dilution, 8 points (20μM-6.67 μM-2.22 μM-740 nM-247 nM-82 nM-27 nM-9 nM) in a finalconcentration of 1% DMSO. When potency of compound series increased,more dilutions were prepared and/or the top concentration was lowered(e.g. 5 μM, 1 μM).

Semi-Quantitative Score:

-   -   #>501 nM    -   ##101-500 nM    -   ###1-100 nM

TABLE IV JAK2 IC₅₀ Values of Compounds Cpd # JAK2 12 ### 14 # 36 ### 37### 57 ### 72 ### 78 ### 92 ## 163 #### 176 ### 197 ### 198 ###

Example 1.3 JAK3 Inhibition Assay

Recombinant human JAK3 catalytic domain (amino acids 781-1124; catalognumber PV3855) was purchased from Invitrogen. 0.025 mU of JAK3 wasincubated with 2.5 μg polyGT substrate (Sigma catalog number P0275) inkinase reaction buffer (25 mM Tris pH 7.5, 0.5 mM EGTA, 0.5 mM Na3VO4, 5mM b-glycerophosphate, 0.01% Triton X-100, 1 μM non-radioactive ATP,0.25 μCi 33P-gamma-ATP (GE Healthcare, catalog number AH9968) finalconcentrations) with or without 5 μL containing test compound or vehicle(DMSO, 1% final concentration), in a total volume of 25 μL, in apolypropylene 96-well plate (Greiner, V-bottom). After 105 min at 30°C., reactions were stopped by adding of 25 μL/well of 150 mM phosphoricacid. All of the terminated kinase reaction was transferred to prewashed(75 mM phosphoric acid) 96 well filter plates (Perkin Elmer catalognumber 6005177) using a cell harvester (Perkin Elmer). Plates werewashed 6 times with 300 μL per well of a 75 mM phosphoric acid solutionand the bottom of the plates was sealed. 40 μL/well of Microscint-20 wasadded, the top of the plates was sealed and readout was performed usingthe Topcount (Perkin Elmer). Kinase activity was calculated bysubtracting counts per minute (cpm) obtained in the presence of apositive control inhibitor (10 μM staurosporine) from cpm obtained inthe presence of vehicle. The ability of a test compound to inhibit thisactivity was determined as:

Percentage inhibition=((cpm determined for sample with test compoundpresent−cpm determined for sample with positive controlinhibitor)divided by(cpm determined in the presence of vehicle cpmdetermined for sample with positive control inhibitor))*100%.

Dose dilution series were prepared for the compounds enabling thetesting of dose-response effects in the JAK3 assay and the calculationof the IC₅₀ for each compound. Each compound was routinely tested atconcentration of 20 μM followed by a ⅓ serial dilution, 8 points (20μM-6.67 μM-2.22 μM-740 nM-247 nM-82 nM-27 nM-9 nM) in a finalconcentration of 1% DMSO. When potency of compound series increased,more dilutions were prepared and/or the top concentration was lowered(e.g. 5 μM, 1 μM).

Semi-Quantitative Score:

-   -   +JAK 3    -   +>501 nM    -   ++101-500 nM    -   +++1-100 nM

TABLE V JAK3 IC₅₀ Values of Compounds Cpd # JAK3 12 +++ 15 ++ 36 ++ 37 +57 + 72 + 78 ++ 163 ++ 176 +++

Example 1.4 TYK2 Inhibition Assay

Recombinant human TYK2 catalytic domain (amino acids 871-1187; catalognumber 08-147) was purchased from Carna biosciences. 5 ng of TYK2 wasincubated with 12.5 μg polyGT substrate (Sigma catalog number P0275) inkinase reaction buffer (25 mM Hepes pH 7.5, 100 mM NaCl, 0.2 mM Na3VO4,0.1% NP-40, 0.1 μM non-radioactive ATP, 0.125 μCi 33P-gamma-ATP (GEHealthcare, catalog number AH9968) final concentrations) with or without5 μL containing test compound or vehicle (DMSO, 1% final concentration),in a total volume of 25 μL, in a polypropylene 96-well plate (Greiner,V-bottom). After 90 min at 30° C., reactions were stopped by adding of25 μL/well of 150 mM phosphoric acid. All of the terminated kinasereaction was transferred to prewashed (75 mM phosphoric acid) 96 wellfilter plates (Perkin Elmer catalog number 6005177) using a cellharvester (Perkin Elmer). Plates were washed 6 times with 300 μL perwell of a 75 mM phosphoric acid solution and the bottom of the plateswas sealed. 40 μL/well of Microscint-20 was added, the top of the plateswas sealed and readout was performed using the Topcount (Perkin Elmer).Kinase activity was calculated by subtracting counts per minute (cpm)obtained in the presence of a positive control inhibitor (10 μMstaurosporine) from cpm obtained in the presence of vehicle. The abilityof a test compound to inhibit this activity was determined as:

Percentage inhibition=((cpm determined for sample with test compoundpresent−cpm determined for sample with positive controlinhibitor)divided by(cpm determined in the presence of vehicle−cpmdetermined for sample with positive control inhibitor))*100%.

Dose dilution series were prepared for the compounds enabling thetesting of dose-response effects in the TYK2 assay and the calculationof the IC₅₀ for each compound. Each compound was routinely tested atconcentration of 20 μM followed by a ⅓ serial dilution, 8 points (20μM-6.67 μM-2.24M-740 nM-247 nM-82 nM-27 nM-9 nM) in a finalconcentration of 1% DMSO. When potency of compound series increased,more dilutions were prepared and/or the top concentration was lowered(e.g. 5 μM, 1 μM).

Semi-Quantitative Score:

-   -   *>1001 nM    -   **501-1000 nM    -   ***101-500 nM    -   ****1-100 nM

TABLE VI TYK2 IC₅₀ Values of Compounds Cpd # TYK2 15 * 36 *** 37 ** 57 *72 * 92 * 163 * 176 ****

Example 2 Cellular Assays Example 2.1 JAK-STAT Signalling Assay

HeLa cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM)containing 10% heat inactivated fetal calf serum, 100 U/mL penicillinand 100 μg/mL streptomycin. HeLa cells were used at 70% confluence fortransfection. 20,000 cells in 87 μL cell culture medium were transientlytransfected with 40 ng pSTAT1(2)-luciferase reporter (Panomics), 8 ng ofLacZ reporter as internal control reporter and 52 ng of pBSK using 0.32μL Jet-PET (Polyplus) as transfection reagent per well in 96-well plateformat. After overnight incubation at 37° C., 10% CO₂, transfectionmedium was removed. 75 μL of DMEM+1.5% heat inactivated fetal calf serumwas added. 15 μL of compound at 6.7× concentration was added for 60 minand then 10 μL of human OSM (Peprotech) at 33 ng/mL final concentration.

All compounds were tested in duplicate starting from 20 μM followed by a⅓ serial dilution, 8 doses in total (20 μM-6.6 μM-2.2 μM-740 nM-250nM-82 nM-27 nM-9 nM) in a final concentration of 0.2% DMSO.

After overnight incubation at 37° C., 10% CO₂ cells were lysed in 100 μLlysis buffer/well (PBS, 0.9 mM CaCl₂, 0.5 mM MgCl₂, 5% Trehalose, 0.025%Tergitol NP9, 0.15% BSA).

40 μL of cell lysate was used to read β-galactosidase activity by adding180 μL βGal solution (30 μl ONPG 4 mg/mL+150 μL β-Galactosidase buffer(0.06 M Na₂HPO₄, 0.04 M NaH₂PO₄, 1 mM MgCl₂)) for 20 min. The reactionwas stopped by addition of 50 μL Na₂CO₃ 1 M. Absorbance was read at 405nm.

Luciferase activity was measured using 40 pt cell lysate plus 40 μl ofSteadylite® as described by the manufacturer (Perkin Elmer), on theEnvision (Perkin Elmer).

10 μM of a pan-JAK inhibitor was used as a positive control (100%inhibition). As negative control 0.5% DMSO (0% inhibition) was used. Thepositive and negative controls were used to calculate z′ and ‘percentinhibition’ (PIN) values.

Percentage inhibition=((fluorescence determined in the presence ofvehicle−fluorescence determined for sample with test compoundpresent)divided by(fluorescence determined in the presence ofvehicle−fluorescence determined for sample without trigger))*100%.

PIN values were plotted for compounds tested in dose-response and EC₅₀values were derived.

TABLE VII JAK-STAT signalling EC₅₀ Values Cpd # EC₅₀ (nM) 12 *** 14 * 15*** 36 *** 37 ** 57 * 72 * 78 * 92 * 163 * 176 **** 182 * 190 * 192 ***197 *** 198 *** * >1001 nM ** 501-1000 nM *** 101-500 nM **** 1-100 nM

Example 2.2 SM/IL-1β Signaling Assay

OSM and IL-1β were shown to synergistically upregulate MMP13 levels inthe human chondrosarcoma cell line SW1353. The cells were seeded in 96well plates at 15,000 cells/well in a volume of 120 μL DMEM (Invitrogen)containing 10% (v/v) FBS and 1% penicillin/streptomycin (InVitrogen)incubated at 37° C. 5% CO₂. Cells were preincubated with 15 μL compoundin M199 medium with 2% DMSO 1 hr before triggering with 15 μL OSM andIL-1β to reach 25 ng/mL OSM and 1 ng/mL IL-1β, and MMP13 levels weremeasured in conditioned medium 48 hours after triggering. MMP13 activitywas measured using an antibody capture activity assay. For this purpose,384 well plates (NUNC, 460518, MaxiSorb black) were coated with 35 μL ofa 1.5 μg/mL anti-human MMP13 antibody (R&D Systems, MAB511) solution for24 hours at 4° C. After washing the wells 2 times with PBS+0.05% Tween,the remaining binding sites were blocked with 100 μL 5% non-fat dry milk(Santa Cruz, sc-2325, Blotto) in PBS for 24 hours at 4° C. Next, thewells were washed 2 times with PBS+0.05% Tween and 35 μL of 1/10dilution of culture supernatant containing MMP13 in 100-fold dilutedblocking buffer was added and incubated for 4 hours at room temperature.Next the wells were washed twice with PBS+0.05% Tween followed by MMP13activation by addition of 35 μL of a 1.5 mM 4-Aminophenylmercuricacetate (APMA) (Sigma, A9563) solution and incubation at 37° C. for 1hour. The wells were washed again with PBS+0.05% Tween and 35 μL MMP13substrate (Biomol, P-126, OmniMMP fluorogenic substrate) was added.After incubation for 24 hours at 37° C. fluorescence of the convertedsubstrate was measured in a Perkin Elmer Wallac EnVision 2102 MultilabelReader (wavelength excitation: 320 nm, wavelength emission: 405 nm).

Percentage inhibition=((fluorescence determined in the presence ofvehicle−fluorescence determined for sample with test compoundpresent)divided by(fluorescence determined in the presence ofvehicle−fluorescence determined for sample without trigger))*100%.

-   -   *>1001 nM    -   **501-1000 nM    -   ***1-500 nM

TABLE VIII MMP13 EC₅₀ Values Cpd # EC₅₀ (nM) 12 * 15 * 36 ** 37 * 57 *72 ** 78 ** 163 * 176 *** 182 * 192 * 197 * 198 *

Example 2.3 PBL Proliferation Assay

Human peripheral blood lymphocytes (PBL) were stimulated with IL-2 andproliferation measured using a BrdU incorporation assay. The PBL werefirst stimulated for 72 hrs with PHA to induce IL-2 receptor, fasted for24 hrs to stop cell proliferation followed by IL-2 stimulation foranother 72 hrs (including 24 hr BrdU labeling). Cells were preincubatedwith test compounds 1 hr before IL-2 addition. Cells were cultured inRPMI 1640 containing 10% (v/v) FBS.

Example 3 In Vivo Models Example 3.1 CIA Model 3.1.1 Materials

Completed Freund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA)were purchased from Difco. Bovine collagen type II (CII),lipopolysaccharide (LPS), and Enbrel were obtained from Chondrex (Isled'Abeau, France); Sigma (P4252, L′Isle d'Abeau, France), Whyett (25 mginjectable syringe, France) Acros Organics (Palo Alto, Calif.),respectively. All other reagents used were of reagent grade and allsolvents were of analytical grade.

3.1.2 Animals

Dark Agouti rats (male, 7-8 weeks old) were obtained from HarlanLaboratories (Maison-Alfort, France). Rats were kept on a 12 hourslight/dark cycle (0700-1900). The temperature was maintained at 22° C.,and food and water were provided ad libitum.

3.1.3 Collagen Induced Arthritis (CIA)

One day before the experiment, CII solution (2 mg/mL) was prepared with0.05 M acetic acid and stored at 4° C. Just before the immunization,equal volumes of adjuvant (IFA) and CII were mixed by a homogenizer in apre-cooled glass bottle in an ice water bath. Extra adjuvant andprolonged homogenization might be required if an emulsion is not formed.0.2 mL of the emulsion was injected intradermally at the base of thetail of each rat on day 1, a second booster intradermal injection (CIIsolution at 2 mg/mL in CFA 0.1 mL saline) was performed on day 9. Thisimmunization method was modified from published methods (Sims et al,2004; Jou et al., 2005).

3.1.4 Study Design

The therapeutic effects of the test compounds were tested in the rat CIAmodel. Rats were randomly divided into equal groups and each groupcontained 10 rats. All rats were immunized on day 1 and boosted on day9. Therapeutic dosing lasted from day 16 to day 30. The negative controlgroup was treated with vehicle (MC 0.5%) and the positive control groupwith Enbrel (10 mg/kg, 3×week., s.c.). A compound of interest wastypically tested at 3 doses, e.g. 3, 10, 30 mg/kg, p.o.

3.1.5 Clinical Assessment of Arthritis

Arthritis was scored according to the method of Khachigian 2006, Lin etal 2007 and Nishida et al. 2004). The swelling of each of the four pawswas ranked with the arthritic score as follows: 0-no symptoms; 1-mild,but definite redness and swelling of one type of joint such as the ankleor wrist, or apparent redness and swelling limited to individual digits,regardless of the number of affected digits; 2-moderate redness andswelling of two or more types of joints; 3-severe redness and swellingof the entire paw including digits; 4-maximally inflamed limb withinvolvement of multiple joints (maximum cumulative clinical arthritisscore 16 per animal) (Nishida et al., 2004).

3.1.6 Change in Body Weight (%) after Onset of Arthritis

Clinically, body weight loss is associated with arthritis (Shelton etal., 2005; Argiles et al., 1998; Rall, 2004; Walsmith et al., 2004)Hence, changes in body weight after onset of arthritis could be used asa non-specific endpoint to evaluate the effect of therapeutics in therat model. The change in body weight (%) after onset of arthritis wascalculated as follows:

${Mice}\text{:}\mspace{20mu} \frac{{{Body}\mspace{14mu} {Weight}_{({{week}\mspace{11mu} 6})}} - {{Body}\mspace{14mu} {Weight}_{({{week}\mspace{11mu} 5})}}}{{Body}\mspace{14mu} {Weight}_{({{week}\mspace{11mu} 5})}} \times 100\%$${Rats}\; \text{:}\mspace{14mu} \frac{{{Body}\mspace{14mu} {Weight}_{({{week}\mspace{11mu} 4})}} - {{Body}\mspace{14mu} {Weight}_{({{week}\mspace{11mu} 3})}}}{{Body}\mspace{14mu} {Weight}_{({{week}\; 3})}} \times 100\%$

3.1.7 Radiology

X-ray photos were taken of the hind paws of each individual animal. Arandom blind identity number was assigned to each of the photos, and theseverity of bone erosion was ranked by two independent scorers with theradiological Larsen's score system as follows: 0-normal with intact bonyoutlines and normal joint space; 1-slight abnormality with any one ortwo of the exterior metatarsal bones showing slight bone erosion;2-definite early abnormality with any three to five of the exteriormetatarsal bones showing bone erosion; 3-medium destructive abnormalitywith all the exterior metatarsal bones as well as any one or two of theinterior metatarsal bones showing definite bone erosions; 4-severedestructive abnormality with all the metatarsal bones showing definitebone erosion and at least one of the inner metatarsal joints completelyeroded leaving some bony joint outlines partly preserved; 5-mutilatingabnormality without bony outlines. This scoring system is a modificationfrom Salvemini et al., 2001; Bush et al., 2002; Sims et al., 2004; Jouet al., 2005.

3.1.8 Histology

After radiological analysis, the hind paws of mice were fixed in 10%phosphate-buffered formalin (pH 7.4), decalcified with rapid bonedecalcifiant for fine histology (Laboratories Eurobio) and embedded inparaffin. To ensure extensive evaluation of the arthritic joints, atleast four serial sections (5 μm thick) were cut and each series ofsections were 100 μm in between. The sections were stained withhematoxylin and eosin (H&E). Histologic examinations for synovialinflammation and bone and cartilage damage were performed double blind.In each paw, four parameters were assessed using a four-point scale. Theparameters were cell infiltration, pannus severity, cartilage erosionand bone erosion. Scoring was performed as follows: 1-normal, 2-mild,3-moderate, 4-marked. These four scores were summed together andrepresented as an additional score, namely the ‘RA total score’.

3.1.9 Micro-Computed Tomography (μCT) Analysis of Calcaneus (Heel Bone):

Bone degradation observed in RA occurs especially at the cortical boneand can be revealed by μCT analysis (Sims N A et al., 2004; Oste L etal., ECTC Montreal 2007). After scanning and 3D volume reconstruction ofthe calcaneus bone, bone degradation was measured as the number ofdiscrete objects present per slide, isolated in silico perpendicular tothe longitudinal axis of the bone. The more the bone that was degraded,the more discrete objects that were measured. 1000 slices, evenlydistributed along the calcaneus (spaced by about 10.8 μm), are analyzed.

3.1.10 Results

Compound 176 was efficacious in all readouts performed in the rat CIAstudy. Statistical significance is obtained for 3 mg/kg in the clinicalscore and paw swelling readouts. Selected additional compounds were alsotested in the rat CIA study, Compound 36 was active at 30 mg/kg,Compound 37 was active at 10 mg/kg.

Example 3.2 Septic Shock Model

Injection of lipopolysaccharide (LPS) induces a rapid release of solubletumour necrosis factor (TNF-alpha) into the periphery. This model isused to analyze prospective blockers of TNF release in vivo.

Six BALB/cJ female mice (20 g) per group were treated at the intendeddosing once, po. Thirty minutes later, LPS (15 μg/kg; E. Coli serotype0111:B4) was injected ip. Ninety minutes later, mice were euthanized andblood was collected. Circulating TNF alpha levels were determined usingcommercially available ELISA kits. Dexamethasone (5 μg/kg) was used as areference anti-inflammatory compound. Selected compounds are tested atone or multiple doses, e.g. 3 and/or 10 and/or 30 mg/kg, po.

Compound 176 exhibited a statistically significant reduction in the TNFrelease (>50%) at 3, and 30 mg/kg po.

Selected additional compounds were also tested in the septic shockmodel, Compound 36 was active at 30 mg/kg, Compounds 37 and 197 wereactive at 3 mg/kg,

Example 3.3 MAB Model

The MAB model allows a rapid assessment of the modulation of an RA-likeinflammatory response by therapeutics (Kachigian L M. Nature Protocols(2006) 2512-2516: Collagen antibody-induced arthritis). DBA/J mice wereinjected i.v. with a cocktail of mAbs directed against collagen II. Oneday later, compound treatment was initiated (vehicle: 10% (v/v) HPPCD).Three days later, mice received an i.p. LPS injection (50 μg/mouse),resulting in a fast onset of inflammation. Compound treatment wascontinued until 10 days after the mAb injection. Inflammation was readby measuring paw swelling and recording the clinical score of each paw.The cumulative clinical arthritis score of four limbs was presented toshow the severity of inflammation. A scoring system is applied to eachlimb using a scale of 0-4, with 4 being the most severe inflammation.

-   -   0 Symptom free    -   1 Mild, but definite redness and swelling of one type of joint        such as the ankle or wrist, or apparent redness and swelling        limited to individual digits, regardless of the number of        affected digits    -   2 Moderate redness and swelling of two or more types of joints    -   3 Severe redness and swelling of the entire paw including digits    -   4 Maximally inflamed limb with involvement of multiple joints

Compound 176, dosed p.o. at 10 and 30 mg/kg reduced the clinical scorewith statistical significance at 30 mg/kg and significantly reducedinflammation at both 10 and 30 mg/kg doses.

Selected additional compounds were also tested in the MAB model,Compounds 36 and 37 were active at 30 mg/kg.

Example 3.4 Oncology Models

In vivo models to validate efficacy of small molecules towards JA K2-driven myeloproliferative diseases are described by Wernig et al.Cancer Cell 13, 311, 2008 and Geron et al. Cancer Cell 13, 321, 2008.

Example 3.5 Mouse IBD Model

In vitro and in vivo models to validate efficacy of small moleculestowards IBD are described by Wirtz et al. 2007.

Example 3.6 Mouse Asthma Model

In vitro and in vivo models to validate efficacy of small moleculestowards asthma are described by Nials et al., 2008; Ip et al. 2006;Pernis et al., 2002; Kudlacz et al., 2008.

Example 4 Toxicity, DMPK and Safety Models Example 4.1 ThermodynamicSolubility

A solution of 1 mg/mL of the test compound is prepared in a 0.2Mphosphate buffer pH7.4 or a 0.1 M citrate buffer pH3.0 at roomtemperature in a glass vial.

The samples are rotated in a Rotator drive STR 4 (Stuart Scientific,Bibby) at speed 3.0 at room temperature for 24 hours.

After 24 hours, 800 μL of the sample is transferred to an eppendorf tubeand centrifuged 5 min at 14000 rpm. 200 μL of the supernatant of thesample is then transferred to a MultiscreenR Solubility Plate(Millipore, MSSLBPC50) and the supernatant is filtered (10-12″ Hg) withthe aid of a vacuum manifold into a clean Greiner polypropylene V-bottom96 well plate (Cat no. 651201). 5 μL of the filtrate is diluted into 95μL (F20) of the same buffer used to incubate in the plate containing thestandard curve (Greiner, Cat no. 651201).

The standard curve for the compound is prepared freshly in DMSO startingfrom a 10 mM DMSO stock solution diluted factor 2 in DMSO (5000 μM) andthen further diluted in DMSO up to 19.5 μM. 3 μL of the dilution seriesas from 5000 μM is then transferred to a 97 μL acetonitrile-buffermixture (50/50). The final concentration range is 2.5 to 150 μM.

The plate is sealed with sealing mats (MA96RD-04S, www.kinesis.co.uk)and samples are measured at room temperature on LCMS (ZQ 1525 fromWaters) under optimized conditions using Quanoptimize to determine theappropriate mass of the molecule.

The samples are analyzed on LCMS with a flow rate of 1 mL/min. Solvent Ais 15 mM ammonia and solvent B is acetonitrile. The sample is run underpositive ion spray on an XBridge C18 3.5 μM (2.1×30 mm) column, fromWaters. The solvent gradient has a total run time of 2 minutes andranges from 5% B to 95% B.

Peak areas are analyzed with the aid of Masslynx software package andpeak areas of the samples are plotted against the standard curve toobtain the solubility of the compound.

Solubility values are reported in μM or μg/mL.

Example 4.2 Aqueous Solubility

Starting from a 10 mM stock in DMSO, a serial dilution of the compoundis prepared in DMSO. The dilution series is transferred to a 96 NUNCMaxisorb plate F-bottom (Cat no. 442404) and 0.2 M phosphate bufferpH7.4 or 0.1 M citrate buffer pH3.0 at room temperature is added.

The final concentration ranged from 200 μM to 2.5 μM in 5 equal dilutionsteps. The final DMSO concentration did not exceed 2%. 200 mM Pyrene isadded to the corner points of each 96 well plate and serves as areference point for calibration of Z-axis on the microscope.

The assay plates are sealed and incubated for 1 hour at 37° C. whileshaking at 230 rpm. The plates are then scanned under a white lightmicroscope, yielding individual pictures of the precipitate perconcentration. The precipitate is analyzed and converted into a numberwhich is plotted onto a graph. The first concentration at which thecompound appears completely dissolved is the concentration reported,however the true concentration lies somewhere between this concentrationand one dilution step higher.

Solubility values are reported in μg/mL.

Example 4.3 Plasma Protein Binding (Equilibrium Dialysis)

A 10 mM stock solution of the compound in DMSO is diluted with a factor5 in DMSO. This solution is further diluted in freshly thawed human,rat, mouse or dog plasma (BioReclamation INC) with a final concentrationof 10 μM and final DMSO concentration of 0.5% (5.5 μl in 1094.5 μlplasma in a PP-Masterblock 96 well (Greiner, Cat no. 780285))

A Pierce Red Device plate with inserts (ThermoScientific, Cat no. 89809)is prepared and filled with 750 μL PBS in the buffer chamber and 500 μLof the spiked plasma in the plasma chamber. The plate is incubated for 4hours at 37° C. while shaking at 230 rpm. After incubation, 120 μL ofboth chambers is transferred to 360 μL acetonitrile in a 96-well roundbottom, PP deep-well plates (Nunc, Cat no. 278743) and sealed with analuminum foil lid. The samples are mixed and placed on ice for 30 min.This plate is then centrifuged 30 min at 1200 rcf at 4° C. and thesupernatant is transferred to a 96 v-bottom PP plate (Greiner, 651201)for analysis on LCMS.

The plate is sealed with scaling mats (MA96RD-04S) of www.kinesis.co.ukand samples are measured at room temperature on LCMS (ZQ 1525 fromWaters) under optimized conditions using Quanoptimize to determine theappropriate mass of the molecule.

The samples are analyzed on LCMS with a flow rate of 1 mL/min. Solvent Awas 15 mM ammonia and solvent B was acetonitrile. The sample was rununder positive ion spray on an XBridge C18 3.5 μM (2.1×30 mm) column,from Waters. The solvent gradient has a total run time of 2 minutes andranges from 5% B to 95% B.

Peak area from the compound in the buffer chamber and the plasma chamberwere considered to be 100% compound. The percentage bound to plasma wasderived from these results and was reported to the LIMS as percentagebound to plasma.

The solubility of the compound in the final test concentration in PBSwas inspected by microscope to indicate whether precipitation isobserved or not.

Example 4.4 Liability for QT Prolongation

Potential for QT prolongation was assessed in the hERG patch clampassay.

4.4.1 Conventional Whole-Cell Patch-Clamp

Whole-cell patch-clamp recordings were performed using an EPC10amplifier controlled by Pulse v8.77 software (HEKA). Series resistancewas typically less than 10 MΩ and compensated by greater than 60%,recordings were not leak subtracted. Electrodes were manufactured fromGC150TF pipette glass (Harvard), resistance was between 2 and 3 MO.

The external bathing solution contained: 135 mM NaCl, 5 mM KCl, 1.8 mMCaCl₂, 5 mM Glucose, 10 mM HEPES, pH 7.4.

The internal patch pipette solution contained: 100 mM Kgluconate, 20 mMKCl, 1 mM CaCl₂, 1 mM MgCl₂, 5 mM Na₂ATP, 2 mM Glutathionc, 11 mM EGTA,10 mM HEPES, pH 7.2.

Drugs were perfused using a Biologic MEV-9/EVH-9 rapid perfusion system.

All recordings were performed on HEK293 cells stably expressing hERGchannels. Cells were cultured on 12 mm round coverslips (German glass,Bellco) anchored in the recording chamber using two platinum rods(Goodfellow). hERG currents were evoked using an activating pulse to +40mV for 1000 ms followed by a tail current pulse to −50 mV for 2000 ms,holding potential was −80 mV. Pulses were applied every 20 s and allexperiments were performed at room temperature.

4.4.2 Data Analysis

IC₅₀ and IC₂₀ values were calculated for each compound tested. The folddifference between the IC₂₀ and the unbound C₁₀ concentrations of thetest compound obtained at relevant therapeutic doses as determined byresults obtained from the rat CIA model was calculated.

For the concentration response curves, peak tail current amplitude wasmeasured during the voltage step to −50 mV. Curve-fitting ofconcentration-response data was performed using the equation:

y=a+[(b−a)/(1+10̂((logc−x)d)]

where a is minimum response, b is maximum response and d is Hill slope,this equation can be used to calculate both IC₅₀ (where y=50 and c isthe IC₅₀ value) and IC₂₀ (where y=20 and c is the IC₂₀ value). GraphPad®Prism® (Graphpad® Software Inc.) software was used for all curvefitting.

Table IX below summarises the results obtained for the selectedcompounds that were tested.

TABLE IX Compound Dose Fold difference 37 10 7 176 3 137 176 10 82

A difference of 100 fold or greater indicates a low potential for QTprolongation. Therefore it can be seen that compared to Compound 37,Compound 176 has a much lower potential QT prolongation issue.

Example 4.5 Microsomal Stability

A 10 mM stock solution of compound in DMSO was diluted 1000 fold in a182 mM phosphate buffer pH7.4 in a 96 deep well plate (Greiner, Catno.780285) and pre-incubated at 37° C.

40 μL of deionised water was added to a well of a polypropylene Matrix2D barcode labelled storage tube (Thermo Scientific) and pre-incubatedat 37° C.

A Glucose-6-phophate-dehydrogenase (G6PDH) working stock solution wasprepared in 182 mM phosphate buffer pH7.4 and placed on ice before use.A co-factor containing MgCl₂, glucose-6-phosphate and NADP+ was preparedin deionised water and placed on ice before use.

A final working solution containing liver microsomes (Xenotech) of aspecies of interest (human, mouse, rat, dog), previously described G6PDHand co-factors was prepared and this mix was incubated for no longerthan 20 minutes at room temperature.

30 μL of the pre-heated compound dilution was added to 40 μL ofpre-heated water in the Matrix tubes and 30 μL of the microsomal mix wasadded. Final reaction concentrations were 3 μM compound, 1 mgmicrosomes, 0.4 U/mL GDPDH, 3.3 mM MgCl₂, 3.3 mM glucose-6-phosphate and1.3 mM NADP-H

To measure percentage remaining of compound at time zero MeOH or ACN wasadded (1:1) to the well before adding the microsomal mix. The plateswere sealed with Matrix Sepra Seals™ (Matrix, Cat. No. 4464) and shakenfor a few seconds ensure complete mixing of all components.

The samples which were not stopped are incubated at 37° C., 300 rpm andafter 1 hour of incubation the reaction was stopped with MeOH or ACN(1:1).

After stopping the reaction the samples were mixed and placed on ice for30 min to precipitate the proteins. The plates were then centrifuged 30min at 1200 rcf at 4° C. and the supernatant was transferred to a 96v-bottom PP plate (Greiner, 651201) for analysis on LCMS.

These plates were sealed with sealing mats (MA96RD-04S) ofwww.kinesis.co.uk and samples were measured at room temperature on LCMS(ZQ 1525 from Waters) under optimized conditions using Quanoptimize todetermine the appropriate mass of the parent molecule.

The samples were analyzed on LCMS with a flow rate of 1 mL/min. SolventA was 15 mM ammonia and solvent B was methanol or acetonitrile,depending on the stop solution used. The samples were run under positiveion spray on an XBridge C18 3.5 μM (2.1×30 mm) column, from Waters. Thesolvent gradient had a total run time of 2 minutes and ranges from 5% Bto 95% B.

Peak area from the parent compound at time 0 was considered to be 100%remaining. The percentage remaining after 1 hour incubation wascalculated from time 0 and was calculated as the percentage remaining.The solubility of the compound in the final test concentration in bufferis inspected by microscope and results are reported.

The data on microsomal stability are expressed as a percentage of thetotal amount of compound remaining after 60 minutes.

TABLE X Microsomal stability Compound # Human (%) Rat (%) 12 70 94 15 4314 36 11.65 72.41 37 70.98 79.47 57 42.75 34.5 72 4.88 2.2 78 78.2193.85 92 30.21 23.6 163 53.64 39.78 176 157.1 90.21 182 19.33 19.23 1902.13 25.64 192 127.2 77.62 197 46.6 59.41 198 65.8 72.45

Example 4.6 Caco2 Permeability

Bi-directional Caco-2 assays were performed as described below. Caco-2cells were obtained from European Collection of Cell Cultures (ECACC,cat 86010202) and used after a 21 day cell culture in 24-well Transwellplates (Fisher TKT-545-020B).

2×10⁵ cells/well were seeded in plating medium consisting ofDMEM+GlutaMAXI+1% NEAA+10% FBS (FetalClone II)+1% Pen/Strep. The mediumwas changed every 2-3 days.

Test and reference compounds (propranolol and rhodaminel 23 orvinblastine, all purchased from Sigma) were prepared in Hanks' BalancedSalt Solution containing 25 mM HEPES (pH7.4) and added to either theapical (125 μL) or basolateral (600 μL) chambers of the Transwell plateassembly at a concentration of 10 μM with a final DMSO concentration of0.25%.

50 μM Lucifer Yellow (Sigma) was added to the donor buffer in all wellsto assess integrity of the cell layers by monitoring Lucifer Yellowpermeation. As Lucifer Yellow (LY) cannot freely permeate lipophilicbarriers, a high degree of LY transport indicates poor integrity of thecell layer.

After a 1 hour incubation at 37° C. while shaking at an orbital shakerat 150 rpm, 70 μL aliquots were taken from both apical (A) and basal (B)chambers and added to 100 μLl 50:50 acetonitrile:water solutioncontaining analytical internal standard carbamazepine) in a 96 wellplate.

Lucifer yellow was measured with a Spectramax Gemini XS (Ex 426 nm andEm 538 nm) in a clean 96 well plate containing 150 μL of liquid frombasolateral and apical side.

Concentrations of compound in the samples were measured by highperformance liquid-chromatography/mass spectroscopy (LC-MS/MS).

Apparent permeability (P_(app)) values were calculated from therelationship:

P _(app)=[compound]_(acceptor final) ×V_(acceptor)/([compound]_(donor initial) ×V _(donor))/T _(ine) ×V_(donor)/Surface area×60×10⁻⁶ cm/s

V=chamber volume

T_(inc)=incubation time.

Surface area=0.33 cm²

The Efflux ratios, as an indication of active efflux from the apicalcell surface, were calculated using the ratio of P_(app) B>A/P_(app)A>B.

The following assay acceptance criteria were used:

Propranolol: P_(app) (A>B) value≧20(×10⁻⁶ cm/s)

Rhodamine 123 or Vinblastine: P_(app) (A>B) value<5 (×10⁻⁶ cm/s) withEfflux ratio ≧5.

Lucifer yellow permeability: ≦100 nm/s

TABLE XI Caco2 Efflux rate P_(app) A > B Compound # (×10−6 cm/sec)Efflux ratio 12 23.3 0.82 15 25.8 0.91 36 36.55 0.89 37 23 0.85 72 14.931.31 78 2 1.5 163 13.4 0.85 176 10.6 0.8 192 12.5 2

Example 4.7 Pharmacokinetic Study in Rodents 4.7.1 Pharmacokinetic Study

Compounds are formulated in PEG200/physiological saline orPEG400/DMSO/physiological saline mixtures for the intravenous route andin 0.5% methylcellulose or 10-30% hydroxylpropyl-β-cyclodextrine pH3 orpH7.4 for the oral route. Test compounds are orally dosed as a singleesophageal gavage at 5-10 mg/kg and intravenously dosed as a bolus viathe caudal vein at 1 mg/kg. Each group consists of 3 rats. Blood samplesare collected either via the jugular vein using cannulated rats or atthe retro-orbital sinus with lithium heparin as anti-coagulant at thetime points in the following range: 0.05 to 8 hours (intravenous route),and 0.25 to 6 or 24 hours (oral route). Whole blood samples arecentrifuged at 5000 rpm for 10 min and the resulting plasma samples arestored at −20° C. pending analysis.

4.7.2 Quantification of Compound Levels in Plasma

Plasma concentrations of each test compound are determined by anLC-MS/MS method in which the mass spectrometer is operated in positiveelectrospray mode.

4.7.3 Determination of Pharmacokinetic Parameters

Pharmacokinetic parameters are calculated using Winnonlin® (Pharsight®,United

Example 4.8 7-Day Rat Toxicity Study

A 7-day oral toxicity study with test compounds was performed inSprague-Dawley male rats to assess their toxic potential andtoxicokinetics, at daily doses of 100, 300 and 500 mg/kg/day, by gavage,at the constant dosage-volume of 5 mL/kg/day.

The test compounds were formulated in 30% (v/v) HPβCD in purified water.Each group included 5 principal male rats as well as 3 satellite animalsfor toxicokinetics. A fourth group was given 30% (v/v) HPβCD in wateronly, at the same frequency, dosage volume and by the same route ofadministration, and acted as the vehicle control group.

The goal of the study was to determine the lowest dose that resulted inno adverse events being identified (no observable adverse effectlevel—NOAEL). Compounds 37 and 176 were tested in this protocol.

The NOAEL levels for Compounds 37 and 176 were both estimated at 500mg/kg.

It will be appreciated by those skilled in the art that the foregoingdescriptions are exemplary and explanatory in nature, an as indicedintended to illustrate the invention and its preferred embodiments.Through routine experimentation, an artisan will recognise apparentmodifications and variations that may be made without departing from thespirit of the invention. Thus, the invention is intended to be definednot by the above description, but by the following claims and theirequivalents.

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All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

From the foregoing description, various modifications and changes in thecompositions and methods of this invention will occur to those skilledin the art. All such modifications coming within the scope of theappended claims are intended to be included therein.

It should be understood that factors such as the differential cellpenetration capacity of the various compounds can contribute todiscrepancies between the activity of the compounds in the in vitrobiochemical and cellular assays.

At least some of the chemical names of compounds of the invention asgiven and set forth in this application, may have been generated on anautomated basis by use of a commercially available chemical namingsoftware program, and have not been independently verified.Representative programs performing this function include the Lexichemnaming tool sold by Open Eye Software, Inc. and the Autonom Softwaretool sold by MDL, Inc. In the instance where the indicated chemical nameand the depicted structure differ, the depicted structure will control.

Chemical structures shown herein were prepared using either ChemDraw® orISIS®/DRAW. Any open valency appearing on a carbon, oxygen or nitrogenatom in the structures herein indicates the presence of a hydrogen atom.Where a chiral center exists in a structure but no specificstereochemistry is shown for the chiral center, both enantiomersassociated with the chiral structure are encompassed by the structure.

1. A compound according to Formula VIa:

or a pharmaceutically acceptable salt thereof.
 2. A pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and apharmaceutically effective amount of a compound according to claim
 1. 3.The pharmaceutical composition of claim 2, wherein the carrier is aparenteral carrier.
 4. The pharmaceutical composition of claim 2,wherein the carrier is an oral carrier.
 5. The pharmaceuticalcomposition of claim 2, wherein the carrier is a topical carrier.
 6. Amethod of treatment, prevention or prophylaxis of diseases involvingcartilage degradation, bone and/or joint degradation, for exampleosteoarthritis; and/or conditions involving inflammation or immuneresponses, such as Crohn's disease, rheumatoid arthritis, psoriasis,allergic airways disease (e.g. asthma, rhinitis), juvenile idiopathicarthritis, colitis, inflammatory bowel diseases, endotoxin-drivendisease states (e.g. complications after bypass surgery or chronicendotoxin states contributing to e.g. chronic cardiac failure), diseasesinvolving impairment of cartilage turnover (e.g. diseases involving theanabolic stimulation of chondrocytes), congenital cartilagemalformations, diseases associated with hypersecretion of IL6 andtransplantation rejection (e.g. organ transplant rejection), comprisingadministering to a subject a therapeutically effective amount of acompound according to claim
 1. 7. A method of treatment, prevention orprophylaxis of proliferative diseases, comprising administering to asubject a therapeutically effective amount of a compound according toclaim
 1. 8. The method according to claim 6, wherein the condition isselected from diseases involving a method of the anabolic stimulation ofchondrocytes.
 9. The method according to claim 6, wherein the conditionis selected from psoriatic arthritis, juvenile rheumatoid arthritis,gouty arthritis, septic or infectious arthritis, reactive arthritis,reflex sympathetic dystrophy, algodystrophy, Tietze syndrome or costalchondritis, fibromyalgia, osteochondritis, neurogenic or neuropathicarthritis, arthropathy, endemic forms of arthritis like osteoarthritisdeformans endemica, Mseleni disease and Handigodu disease; degenerationresulting from fibromyalgia, systemic lupus erythematosus, sclerodermaand ankylosing spondylitis.
 10. A method of treatment or prophylaxis ofa disease involving degradation of cartilage, comprising administeringto a subject a therapeutically effective amount of a compound accordingto claim
 1. 11. A method of treatment or prevention of osteoarthritis,comprising administering to a subject, a therapeutically effectiveamount of a compound according to claim
 1. 12. A method of treatment orprevention of a condition or a disease involving inflammation,comprising administering to a subject a therapeutically effective amountof a compound according to claim
 1. 13. A method of treatment,prevention or prophylaxis of diseases involving cartilage degradation,bone and/or joint degradation, for example osteoarthritis; and/orconditions involving inflammation or immune responses, such as Crohn'sdisease, rheumatoid arthritis, psoriasis, allergic airways disease (e.g.asthma, rhinitis), juvenile idiopathic arthritis, colitis, inflammatorybowel diseases, endotoxin-driven disease states (e.g. complicationsafter bypass surgery or chronic endotoxin states contributing to e.g.chronic cardiac failure), diseases involving impairment of cartilageturnover (e.g. diseases involving the anabolic stimulation ofchondrocytes), congenital cartilage malformations, diseases associatedwith hypersecretion of IL6 and transplantation rejection (e.g. organtransplant rejection), comprising administering to a subject atherapeutically effective amount of a pharmaceutical compositionaccording to claim
 2. 14. The method according to claim 13, wherein thecondition is selected from diseases involving a method of the anabolicstimulation of chondrocytes.
 15. The method according to claim 13,wherein the condition is selected from psoriatic arthritis, juvenilerheumatoid arthritis, gouty arthritis, septic or infectious arthritis,reactive arthritis, reflex sympathetic dystrophy, algodystrophy, Tietzesyndrome or costal chondritis, fibromyalgia, osteochondritis, neurogenicor neuropathic arthritis, arthropathy, endemic forms of arthritis likeosteoarthritis deformans endemica, Mseleni disease and Handigodudisease; degeneration resulting from fibromyalgia, systemic lupuserythematosus, scleroderma and ankylosing spondylitis.
 16. A method oftreatment, prevention or prophylaxis of proliferative diseases,comprising administering to a subject a therapeutically effective amountof a pharmaceutical composition according to claim
 2. 17. A method oftreatment or prophylaxis of a disease involving degradation ofcartilage, comprising administering to a subject a therapeuticallyeffective amount of a pharmaceutical composition according to claim 2.18. A method of treatment or prevention of osteoarthritis, comprisingadministering to a subject, a therapeutically effective amount of apharmaceutical composition according to claim
 2. 19. A method oftreatment or prevention of a condition or a disease involvinginflammation, comprising administering to a subject a therapeuticallyeffective amount of a pharmaceutical composition according to claim 2.